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3 Swab the top of the culture bottle and reincu- Note: Recognition of bacteria in a Gram smear,
bate. especially Gram negative organisms, is often
difficult due to background debris.
Contamination of blood cultures
Contamination can occur when an aseptic technique Bacteria that may be found in contaminated bank
is not used at the time the blood is collected or at a blood
later stage in the laboratory when subculturing. These are usually organisms that are capable of
Frequent contaminants of blood cultures include growing at room temperature or below. They
commensal staphylococci, micrococci, and diph- include coliforms, Achromobacter species, and
theroids or contaminants from the environment pseudomonads and less commonly Yersinia species.
such as species of Bacillus or Acinetobacter.
Occasionally in immunocompromised patients,
organisms usually considered ‘contaminants’ may be
pathogenic, especially fungi.
Contamination is indicated when an organism is
7.15 Examination of semen
recovered from only one bottle when it should have
grown in both thioglycollate broth and the diphasic Analysis of semen (seminal fluid) is included in this chapter
culture medium or when a mixed microbial flora is because it is often performed in a microbiology laboratory.
isolated. The values and clinical notes contained in this subunit have
Note: An obligatory anaerobe will be isolated from the thio- been referenced from the WHO laboratory manual for the
glycollate culture only. examination of human semen and semen-cervical mucus inter-
action (see Further information)
7.14–7.15
M ICROBIOLOGICAL TESTS 131
be kept as near as possible to body temperature. Normal motility: Over 50% of spermatozoa are
This is best achieved by placing the container motile within 60 minutes of ejaculation.
inside a plastic bag and transporting it in a The spermatozoa remain motile for several hours.
pocket in the person’s clothing.
When more than 60% of spermatozoa are non-
motile, examine an eosin preparation to assess
LABORATORY EXAMINATION OF SEMEN whether the spermatozoa are viable or non-viable
Caution: Handle semen with care because it may (see following text).
contain infectious pathogens, e.g. HIV, hepatitis Presence of cells in semen: Report when more than a few leu-
viruses, herpes viruses. cocytes (pus cells) or red cells are present. When pus cells are
seen, examine a Gram stained smear for bacteria.
– Focus the specimen using the 10 objective. – Using the 10 objective with the condenser iris
Close the condenser iris sufficiently to give good closed sufficiently to give good contrast, count the
contrast. Ensure the spermatozoa are evenly dis- number of spermatozoa in an area of 2 sq mm,
tributed (if not, re-mix the semen and examine a i.e. 2 large squares.
new preparation). Note: The total area of an Improved Neubauer and a
Bürker ruled chamber is 9 sq mm, i.e. 9 large squares.
7.15
132 DISTRICT LABORATORY PRACTICE IN TROPICAL COUNTRIES
7.15–7.16