PATIENT INSTRUCTIONS FOR SAMPLE COLLECTION, SAFE
HANDLING AND DELIVERY
1. Patient should be given clear and simple instructions explaining the need for semen analysis and what is required for specimen collection.
2. Patient should be informed about the importance of abstinence time. Ejaculate must be collected after 3-5 days (but not more than 7 days) of abstinence.
3. Samples should be obtained by masturbation and collected in a warm (20-40ºC), sterile, nontoxic plastic or glass wide-mouth container. Prior to sample collection, the patient must void and wash hands and genitals to minimize the chances of contamination.
4. Use of lubricants and saliva should be avoided as their potential toxicity might influence the result. Semen samples should be protected from extremes of temperature (<20ºC or >40ºC) during transport to the laboratory.
5. All sample containers are labeled with adequate information to eliminate any chances of error.
6. Regular condoms should not be used because of their spermicidal effect. Ideally, the samples must be collected close to the laboratory. If the specimen cannot be produced close to the laboratory, it must be delivered to the laboratory as soon as possible, certainly within 1 hour of collection. During this period, the sample has to be kept warm by carrying it next to the body, and temperature extremes must be avoided.
Macroscopic Examination
Semen Age
Record the time when the sample was received, to the liquefaction time.
Liquefaction
Incubation of the sample must be carried out at either ambient temperature or by placing the specimen in an incubator at 37ºC. A normal sample usually liquefies within 60 minutes at room temperature, although usually this occurs within 15-20 minutes. It is determined by the time required for the gelatinous mass to liquefy. A normal sample might contain gel-like gelatinous corpuscles that do not liquefy. Exact liquefaction time is of no diagnostic importance unless >2 hours elapse without any change. This may indicate poor prostatic secretion since the liquefying enzymes are derived from the prostate gland. On the other hand, absence of coagulation may indicate ejaculatory duct obstruction or congenital absence of seminal vesicles. For samples that do not liquefy, the sample can be mixed by rapidly mixing with a regular transfer pipette or, if the sample is viscous, viscosity can be broken by using a viscosity treatment system.
Color and Odor
It is important to note the color. Normal semen is homogeneously opaque, whitish grey or pearly white. The semen odor is unmistakable and pungent because of sperm oxidation. A yellowish tinge to the semen appears with an increase in the days of abstinence or probably due to carotene pigment. More pronounced yellow discoloration may indicate jaundice or contamination of semen with urine (e.g. bladder neck dysfunction). Drugs like methylene blue and pyridium may also color the semen. Fresh blood (hematospermia) will give semen a reddish tinge, while old blood gives it a brownish tinge. This could be due to the presence of inflammation. Prostatic secretions give semen a strong distinctive odor. Absence or uncharacteristic odor could be associated with an infection.
Volume
Volume of the ejaculate should be measured by transferring the liquefied sample into a graduated 15 mL conical centrifuge tube. The normal volume of ejaculate after 2-5 days of sexual abstinence is about 2-6 mL. Retrograde ejaculation, obstruction of lower urinary tract (urethra, congenital absence
of vas deferens, seminal vesicles) may yield low volume. According to the WHO laboratory manual, the reference value for semen volume is ≥2.0mL; however, for clinical purposes; semen volume is differentiated into three categories to facilitate interpretation and diagnosis:
Aspermia: No semen produced after orgasm (seen in certain clinical conditions).
Hypospermia: <0.5 mL of semen ejaculated (partial or complete retrograde flow of semen, accessory glands impairment).
Hyperspermia: > 6 mL of semen ejaculated (long period of sexual abstinence or overproduction of fluids from the accessory sex glands).
If the volume is <1 mL it is important to determine if the sample is complete. The highest sperm concentration is seen in the initial ejaculate.
Viscosity
Viscosity measures the seminal fluid’s resistance to flowing. It is measured by the length of the ‘thread-lines’ or ‘spinnbarkeit.’ It can be estimated by using a glass rod and observing the length of thread that forms on withdrawal of the rod. A normal sample leaves small, discrete drops; abnormal samples will form threads more than 2 cm long. High viscosity may interfere with determinations of sperm motility, concentration and antibody coating of spermatozoa. Viscosity can be categorized as ‘normal’, ‘moderate’ or ‘high’. Viscous samples can be treated by a viscosity treatment system containing a premeasured vial of chymotrypsin (5 mg/vial (Conception Technologies,San Diego, CA). The sample can be swirled and left in the incubator for another 10-15 min till viscosity is completely broken down and the sample is suitable for analysis.
pH
The pH of liquefied semen is determined by using pH test strips; pH 6.5 to 10 has been found most suitable for this purpose. A drop of semen is spread evenly onto the pH paper. After 30 seconds, the color of the impregnated zone is compared with the calibrated strip. Normal semen pH is in the range of 7.2 to 8.2, and it does tend to increase with time after ejaculation. Any change in the normal range of pH may be caused by inflammation of the prostate or seminal vesicles.
MICROSCOPIC EXAMINATION
Wet Preparation Examination
Load a 5 μL of well-mixed semen on a clean, warmed microscope slide with a cover slip on top (18 × 18 mm). If a 22 × 22 mm cover slip is used, the semen volume on the microscope slide should be 10 μL). This preparation has a depth of approximately ~20 μm. A depth less than 20 μm will hamper the rotational movements of the spermatozoa. Care should be taken to avoid formation of air bubbles that can be trapped between the cover slip and the slide. It is important to wait for the drifting to cease/stabilize before examination. In addition, a variety of other disposable two-well 20 μm
counting chambers are also available. A phase contrast microscope is recommended for all examinations of unstained preparations of fresh/washed semen. Initial examination is done under 10× objective, which provides an overview for determining mucus strands, sperm aggregation, and evenness of spread of spermatozoa on the slide. Subsequently, the sample should be examined for count and motility under 40× objective.
Sperm Concentration
Determining accurate sperm concentration (million/mL of ejaculate) and total sperm count (million sperm per ejaculate) is important. The most accurate method of determining sperm concentration is volumetric dilution and hemocytometry. Gently mixing the semen sample using a positive displacement pipette before the volume is withdrawn is essential for an accurate determination of sperm concentration.
Hemocytometry
Principle
A fixed volume of a liquefied semen aliquot is used and fixed sperm are counted in a Neubauer hemocytometer chamber. Dilution of 1:19 is usually employed. Dilutions may be made in small, clean, glass or plastic vials. Extreme care must be taken while making dilutions and preparing the
hemocytometer.
Reagents
1. The diluent consists of 50 g sodium bicarbonate
2. 10 ml of 35% of formaldehyde solution, and 0.25 g trypan blue dissolved in reagent water up to 1 liter.
Procedure
1. Filter the solution through Whatman No.1 papers into a clean bottle and store it at 4ºC.
2. Add 50 μL liquefied semen to 950 μL diluent. Use a positive displacement pipette to ensure accurate handling of the viscous semen.
3. These dilutions can be stored for up to 4 weeks at 4ºC.
4. Place the hemocytometer cover slip over the chamber.
5. Vortex the diluent for 10 s. Transfer 10 μL to each chamber.
6. Leave the hemocytometer in a humid chamber for 10 to 15 minutes for the spermatozoa to settle down onto the counting grid.
7. Count the spermatozoa using a 20× objective phase-contrast optics. The central square of the grid in an improved Neubauer chamber contains 25 large squares, each containing 16 small squares. The number of squares counted depends on the number of spermatozoa seen in the first large square as follows:
i. < 10 spermatozoa in the first large square - count the whole grid of 25 large squares;.
ii.10 - 40 spermatozoa per square - count only 10 large squares (two horizontal or vertical rows); and
iii. >40 spermatozoa per square - count spermatozoa in the five large squares (the four corners plus the center).
Results
1. Counts of two hemocytometer chambers should be within 5% of their average. If not, discard, remix sample, and prepare another sample to be loaded on the hemocytometer, i.e. (higher value - lower value) must be < (sum of values/20) for the counts to be acceptable
2. Sperm concentration (10ˆ6/mL) = total number of spermatozoa/appropriate correction factor
Total sperm count = Sperm concentration × ejaculate volume.
Sperm Motility Assessment
Sperm motility is the ratio of the number of motile sperm to total number of sperm in a given volume and is expressed as a percentage. Several scoring systems exist for sperm motility assessments, but a simple grading system is recommended. This provides an assessment of sperm motility without requiring sophisticated equipments. According to the WHO laboratory manual (WHO, 1999), five microscopic fields are assessed in a systematic way to classify 200 spermatozoa. The motility of each spermatozoon is graded into one of four groups:
a. Rapid progressive motility (i.e. > 25 μm/s at 37 ºC and > 20 μm/s at 20ºC; note that 25 μm is approximately equal to 5 head lengths or half a tail length).
b. Slow or sluggish progressive motility
c. Non-progressive motility (< 5 μm/s)
d. Immotility
A normal semen analysis must contain at least 50% progressively motile spermatozoa.
Evaluation of Morphology Assessment
For a complete evaluation of a semen sample, the assessment of the morphological characteristics of the spermatozoa is important. The staining of a seminal smear allows the quantitative evaluation of normal and abnormal sperm forms in an ejaculate.
Smear Preparation
Slides should be pre-cleaned with 95% ethanol to allow firm attachment of smears. A small drop of semen, approximately a 5μL aliquot, is placed on the slide. The fraction is then pulled out into a smear with a second slide; this is called the ‘feathering’ technique. This is done with minimum force to ensure that the spermatozoa tails do not fall apart, and care is taken to guarantee that the smear is not too thick. Two smears are made from each sample. If the sperm concentration is > 20 × 10ˆ6/ mL, then 5 μL of semen can be used; if the sperm concentration is < 20 × 10ˆ6/ mL, then 10-20 μL of semen is used. Smears are air-dried and fixed in 95% ethanol for 15 minutes.
Staining Methods
The numerous staining techniques available include as the Papanicolaou, Giemsa, Shorr, modified Bryan-Leishman and Diff-Quik methods with Papanicolaou and Diff-Quik being the more common.
Normal Reference Values of Semen Variables
Each laboratory must determine its own reference range for each variable. According to the World Health Organization guidelines (WHO, 1999) the following reference values for the semen sample are suggested:
•Volume 2.0 ml or more
•pH 7.2 or more
•Sperm concentration 20x10ˆ6spermatozoa/ml or more
•Total sperm count 40x10ˆ6spermatozoa or more
•Motility 50% or more motile (grade a+b) or 25% or more with progressive motility (grade a) within 60 min aftercollection.
•Vitality 75% or more live
•White blood cells Fewer than 1x10ˆ6/ml.
DEFINITIONS OF SEMEN CLASSIFICATIONS
-Normozoospermia When all the spermatozoal parameters are normal together with normal seminal plasma and WBCs, and there is no agglutination.
-Oligozoospermia When sperm concentration is <20million/ml.
-Asthenozoospermia Fewer than 50% spermatozoa with forward progression (categories (a) and (b) or fewer than 25% spermatozoa with category (a) movement).
-Teratozoospermia spermatozoa with decreased % of normal morphology.
-Oligoasthenoterato/ signifies disturbance of all the three zoospermia variables
-Azoospermia No spermatozoa in the ejaculate
-Aspermia -no ejaculate
SEMEN MICROBIOLOGY
Infection of the male reproductive tract can directly or indirectly cause infertility. Inflammation caused by infection or various disorders can affect the secretory function of both the prostate and seminal vesicles. Asymptomatic infections of the prostate can cause partial or complete obstruction of the ejaculatory duct resulting in oligospermia and even azoospermia. Infection of the seminal vesicles often causes substantial reduction in ejaculate volume and a low seminal fructose concentration. Microbiological examination of the semen is required to differentiate a specific microbiological-induced pyospermia from other abnormalities that causes an increase in leukocytes.
Pyospermia
Pyospermia is a laboratory finding categorized as the abnormal presence of leukocytes in human ejaculate. Pyospermia is established when the concentration of seminal WBCs is in the range between 5 × 10ˆ5/mL and 5× 10ˆ6/mL seminal fluid during semen analysis. Numerous studies have demonstrated that leukocytes in ejaculate have a physiological effect on sperm function, which may further impact male infertility (Wolff H, Anderson DJ, 1988). The differential diagnosis of symptomatic pyospermia includes infection, autoimmune disease, and inflammation of the accessory sex glands and the lower male urogenital tract. Urogenital infections include acute and chronic prostatitis, seminal vesiculitis, epididymo-orchitis, cystitis, urethritis, urethral stricture, stone disease, foreign bodies, upper urinary tract infection, retrograde ejaculation, and localized sepsis of the adjacent lower gastrointestinal tract and asymptomatic bacteriuria. Chronic infections that may result in pyospermia include fungal, mycobacterial, and congenital lesions that cause urogenital tract infection.
Organisms Found in Semen
Many organisms found in semen are actually contaminants from the patient’s skin or from the air at the time of collection. Not all are associated with pyospermia, hence caution must be exercised in interpretation of positive cultures.
HANDLING AND DELIVERY
1. Patient should be given clear and simple instructions explaining the need for semen analysis and what is required for specimen collection.
2. Patient should be informed about the importance of abstinence time. Ejaculate must be collected after 3-5 days (but not more than 7 days) of abstinence.
3. Samples should be obtained by masturbation and collected in a warm (20-40ºC), sterile, nontoxic plastic or glass wide-mouth container. Prior to sample collection, the patient must void and wash hands and genitals to minimize the chances of contamination.
4. Use of lubricants and saliva should be avoided as their potential toxicity might influence the result. Semen samples should be protected from extremes of temperature (<20ºC or >40ºC) during transport to the laboratory.
5. All sample containers are labeled with adequate information to eliminate any chances of error.
6. Regular condoms should not be used because of their spermicidal effect. Ideally, the samples must be collected close to the laboratory. If the specimen cannot be produced close to the laboratory, it must be delivered to the laboratory as soon as possible, certainly within 1 hour of collection. During this period, the sample has to be kept warm by carrying it next to the body, and temperature extremes must be avoided.
Macroscopic Examination
Semen Age
Record the time when the sample was received, to the liquefaction time.
Liquefaction
Incubation of the sample must be carried out at either ambient temperature or by placing the specimen in an incubator at 37ºC. A normal sample usually liquefies within 60 minutes at room temperature, although usually this occurs within 15-20 minutes. It is determined by the time required for the gelatinous mass to liquefy. A normal sample might contain gel-like gelatinous corpuscles that do not liquefy. Exact liquefaction time is of no diagnostic importance unless >2 hours elapse without any change. This may indicate poor prostatic secretion since the liquefying enzymes are derived from the prostate gland. On the other hand, absence of coagulation may indicate ejaculatory duct obstruction or congenital absence of seminal vesicles. For samples that do not liquefy, the sample can be mixed by rapidly mixing with a regular transfer pipette or, if the sample is viscous, viscosity can be broken by using a viscosity treatment system.
Color and Odor
It is important to note the color. Normal semen is homogeneously opaque, whitish grey or pearly white. The semen odor is unmistakable and pungent because of sperm oxidation. A yellowish tinge to the semen appears with an increase in the days of abstinence or probably due to carotene pigment. More pronounced yellow discoloration may indicate jaundice or contamination of semen with urine (e.g. bladder neck dysfunction). Drugs like methylene blue and pyridium may also color the semen. Fresh blood (hematospermia) will give semen a reddish tinge, while old blood gives it a brownish tinge. This could be due to the presence of inflammation. Prostatic secretions give semen a strong distinctive odor. Absence or uncharacteristic odor could be associated with an infection.
Volume
Volume of the ejaculate should be measured by transferring the liquefied sample into a graduated 15 mL conical centrifuge tube. The normal volume of ejaculate after 2-5 days of sexual abstinence is about 2-6 mL. Retrograde ejaculation, obstruction of lower urinary tract (urethra, congenital absence
of vas deferens, seminal vesicles) may yield low volume. According to the WHO laboratory manual, the reference value for semen volume is ≥2.0mL; however, for clinical purposes; semen volume is differentiated into three categories to facilitate interpretation and diagnosis:
Aspermia: No semen produced after orgasm (seen in certain clinical conditions).
Hypospermia: <0.5 mL of semen ejaculated (partial or complete retrograde flow of semen, accessory glands impairment).
Hyperspermia: > 6 mL of semen ejaculated (long period of sexual abstinence or overproduction of fluids from the accessory sex glands).
If the volume is <1 mL it is important to determine if the sample is complete. The highest sperm concentration is seen in the initial ejaculate.
Viscosity
Viscosity measures the seminal fluid’s resistance to flowing. It is measured by the length of the ‘thread-lines’ or ‘spinnbarkeit.’ It can be estimated by using a glass rod and observing the length of thread that forms on withdrawal of the rod. A normal sample leaves small, discrete drops; abnormal samples will form threads more than 2 cm long. High viscosity may interfere with determinations of sperm motility, concentration and antibody coating of spermatozoa. Viscosity can be categorized as ‘normal’, ‘moderate’ or ‘high’. Viscous samples can be treated by a viscosity treatment system containing a premeasured vial of chymotrypsin (5 mg/vial (Conception Technologies,San Diego, CA). The sample can be swirled and left in the incubator for another 10-15 min till viscosity is completely broken down and the sample is suitable for analysis.
pH
The pH of liquefied semen is determined by using pH test strips; pH 6.5 to 10 has been found most suitable for this purpose. A drop of semen is spread evenly onto the pH paper. After 30 seconds, the color of the impregnated zone is compared with the calibrated strip. Normal semen pH is in the range of 7.2 to 8.2, and it does tend to increase with time after ejaculation. Any change in the normal range of pH may be caused by inflammation of the prostate or seminal vesicles.
MICROSCOPIC EXAMINATION
Wet Preparation Examination
Load a 5 μL of well-mixed semen on a clean, warmed microscope slide with a cover slip on top (18 × 18 mm). If a 22 × 22 mm cover slip is used, the semen volume on the microscope slide should be 10 μL). This preparation has a depth of approximately ~20 μm. A depth less than 20 μm will hamper the rotational movements of the spermatozoa. Care should be taken to avoid formation of air bubbles that can be trapped between the cover slip and the slide. It is important to wait for the drifting to cease/stabilize before examination. In addition, a variety of other disposable two-well 20 μm
counting chambers are also available. A phase contrast microscope is recommended for all examinations of unstained preparations of fresh/washed semen. Initial examination is done under 10× objective, which provides an overview for determining mucus strands, sperm aggregation, and evenness of spread of spermatozoa on the slide. Subsequently, the sample should be examined for count and motility under 40× objective.
Sperm Concentration
Determining accurate sperm concentration (million/mL of ejaculate) and total sperm count (million sperm per ejaculate) is important. The most accurate method of determining sperm concentration is volumetric dilution and hemocytometry. Gently mixing the semen sample using a positive displacement pipette before the volume is withdrawn is essential for an accurate determination of sperm concentration.
Hemocytometry
Principle
A fixed volume of a liquefied semen aliquot is used and fixed sperm are counted in a Neubauer hemocytometer chamber. Dilution of 1:19 is usually employed. Dilutions may be made in small, clean, glass or plastic vials. Extreme care must be taken while making dilutions and preparing the
hemocytometer.
Reagents
1. The diluent consists of 50 g sodium bicarbonate
2. 10 ml of 35% of formaldehyde solution, and 0.25 g trypan blue dissolved in reagent water up to 1 liter.
Procedure
1. Filter the solution through Whatman No.1 papers into a clean bottle and store it at 4ºC.
2. Add 50 μL liquefied semen to 950 μL diluent. Use a positive displacement pipette to ensure accurate handling of the viscous semen.
3. These dilutions can be stored for up to 4 weeks at 4ºC.
4. Place the hemocytometer cover slip over the chamber.
5. Vortex the diluent for 10 s. Transfer 10 μL to each chamber.
6. Leave the hemocytometer in a humid chamber for 10 to 15 minutes for the spermatozoa to settle down onto the counting grid.
7. Count the spermatozoa using a 20× objective phase-contrast optics. The central square of the grid in an improved Neubauer chamber contains 25 large squares, each containing 16 small squares. The number of squares counted depends on the number of spermatozoa seen in the first large square as follows:
i. < 10 spermatozoa in the first large square - count the whole grid of 25 large squares;.
ii.10 - 40 spermatozoa per square - count only 10 large squares (two horizontal or vertical rows); and
iii. >40 spermatozoa per square - count spermatozoa in the five large squares (the four corners plus the center).
Results
1. Counts of two hemocytometer chambers should be within 5% of their average. If not, discard, remix sample, and prepare another sample to be loaded on the hemocytometer, i.e. (higher value - lower value) must be < (sum of values/20) for the counts to be acceptable
2. Sperm concentration (10ˆ6/mL) = total number of spermatozoa/appropriate correction factor
Total sperm count = Sperm concentration × ejaculate volume.
Sperm Motility Assessment
Sperm motility is the ratio of the number of motile sperm to total number of sperm in a given volume and is expressed as a percentage. Several scoring systems exist for sperm motility assessments, but a simple grading system is recommended. This provides an assessment of sperm motility without requiring sophisticated equipments. According to the WHO laboratory manual (WHO, 1999), five microscopic fields are assessed in a systematic way to classify 200 spermatozoa. The motility of each spermatozoon is graded into one of four groups:
a. Rapid progressive motility (i.e. > 25 μm/s at 37 ºC and > 20 μm/s at 20ºC; note that 25 μm is approximately equal to 5 head lengths or half a tail length).
b. Slow or sluggish progressive motility
c. Non-progressive motility (< 5 μm/s)
d. Immotility
A normal semen analysis must contain at least 50% progressively motile spermatozoa.
Evaluation of Morphology Assessment
For a complete evaluation of a semen sample, the assessment of the morphological characteristics of the spermatozoa is important. The staining of a seminal smear allows the quantitative evaluation of normal and abnormal sperm forms in an ejaculate.
Smear Preparation
Slides should be pre-cleaned with 95% ethanol to allow firm attachment of smears. A small drop of semen, approximately a 5μL aliquot, is placed on the slide. The fraction is then pulled out into a smear with a second slide; this is called the ‘feathering’ technique. This is done with minimum force to ensure that the spermatozoa tails do not fall apart, and care is taken to guarantee that the smear is not too thick. Two smears are made from each sample. If the sperm concentration is > 20 × 10ˆ6/ mL, then 5 μL of semen can be used; if the sperm concentration is < 20 × 10ˆ6/ mL, then 10-20 μL of semen is used. Smears are air-dried and fixed in 95% ethanol for 15 minutes.
Staining Methods
The numerous staining techniques available include as the Papanicolaou, Giemsa, Shorr, modified Bryan-Leishman and Diff-Quik methods with Papanicolaou and Diff-Quik being the more common.
Normal Reference Values of Semen Variables
Each laboratory must determine its own reference range for each variable. According to the World Health Organization guidelines (WHO, 1999) the following reference values for the semen sample are suggested:
•Volume 2.0 ml or more
•pH 7.2 or more
•Sperm concentration 20x10ˆ6spermatozoa/ml or more
•Total sperm count 40x10ˆ6spermatozoa or more
•Motility 50% or more motile (grade a+b) or 25% or more with progressive motility (grade a) within 60 min aftercollection.
•Vitality 75% or more live
•White blood cells Fewer than 1x10ˆ6/ml.
DEFINITIONS OF SEMEN CLASSIFICATIONS
-Normozoospermia When all the spermatozoal parameters are normal together with normal seminal plasma and WBCs, and there is no agglutination.
-Oligozoospermia When sperm concentration is <20million/ml.
-Asthenozoospermia Fewer than 50% spermatozoa with forward progression (categories (a) and (b) or fewer than 25% spermatozoa with category (a) movement).
-Teratozoospermia spermatozoa with decreased % of normal morphology.
-Oligoasthenoterato/ signifies disturbance of all the three zoospermia variables
-Azoospermia No spermatozoa in the ejaculate
-Aspermia -no ejaculate
SEMEN MICROBIOLOGY
Infection of the male reproductive tract can directly or indirectly cause infertility. Inflammation caused by infection or various disorders can affect the secretory function of both the prostate and seminal vesicles. Asymptomatic infections of the prostate can cause partial or complete obstruction of the ejaculatory duct resulting in oligospermia and even azoospermia. Infection of the seminal vesicles often causes substantial reduction in ejaculate volume and a low seminal fructose concentration. Microbiological examination of the semen is required to differentiate a specific microbiological-induced pyospermia from other abnormalities that causes an increase in leukocytes.
Pyospermia
Pyospermia is a laboratory finding categorized as the abnormal presence of leukocytes in human ejaculate. Pyospermia is established when the concentration of seminal WBCs is in the range between 5 × 10ˆ5/mL and 5× 10ˆ6/mL seminal fluid during semen analysis. Numerous studies have demonstrated that leukocytes in ejaculate have a physiological effect on sperm function, which may further impact male infertility (Wolff H, Anderson DJ, 1988). The differential diagnosis of symptomatic pyospermia includes infection, autoimmune disease, and inflammation of the accessory sex glands and the lower male urogenital tract. Urogenital infections include acute and chronic prostatitis, seminal vesiculitis, epididymo-orchitis, cystitis, urethritis, urethral stricture, stone disease, foreign bodies, upper urinary tract infection, retrograde ejaculation, and localized sepsis of the adjacent lower gastrointestinal tract and asymptomatic bacteriuria. Chronic infections that may result in pyospermia include fungal, mycobacterial, and congenital lesions that cause urogenital tract infection.
Organisms Found in Semen
Many organisms found in semen are actually contaminants from the patient’s skin or from the air at the time of collection. Not all are associated with pyospermia, hence caution must be exercised in interpretation of positive cultures.
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