c. Specimens for streptococcus tjping 
Ver\’ few laboratories are in a position to 
type streptococci. If isolates are to be sent 
to the NCDC for typing, they should be sub- 
mitted only after consultation with the Strep- 
tococcus Unit of the Laboratory Program 
and then, as pure cultures on blood agar 
slants. 
8. Haemophilus Injections of the Pharynx or 
Conjunctiva 
Specimens for the isolation of the agent 
Blood, sputum, throat, or nasopharyngeal swabs, 
and purulent exudates are collected in the 
customary manner, using aseptic technic where 
feasible. Since use of swabs made of wooden 
sticks or wire inhibit the growth of these organ- 
isms, small swabs made of cotton attached to 
some inert, synthetic material such as nylon or 
Teflon, which is not inhibitory, should be used. 
Teflon tubing has proved to be satisfactory for 
this purpose. A small, sterile, dry swab may be 
rotated gently over the conjunctiva or pharyn- 
geal mucosa to obtain some of the exudate if 
any is present. The swab should be immediately 
placed in a tube of semisolid agar for transport 
to the laboratory where it is incubated for about 
4 hours. Plates of transparent agar and blood 
are then streaked for isolation of the organism. 
Blood should be inoculated into a flask of broth 
with a large surface area. Sputum and throat 
specimens are cultured on one of the transparent 
agars as well as on blood agar. The swab should 
not be left in the tube of semisolid agar during 
incubation. 
9. Leptospirosis. This disease should be considered 
in all cases of febrile illness of unknown origin 
and it may suggest aseptic meningitis, or non- 
paralytic ]X)liomyelitis. Rats, dogs, cattle, swine, 
and many wild animals are common animal 
reservoirs. Infection is transmitted to man by 
urine containing the agent and occurs when 
leptospires enter the body through the mucous 
membranes of the mouth, nose, throat, eyes, 
lungs, or abraded skin. 
a. .Specimen.s for the isolation of the agent 
Blood, urine, and cerebrospinal fluid may be 
collected for culture and/or animal inocula- 
tion with a view to recovery of the organism. 
Blood taken during the febrile stage of 
illness is the most reliable for culture, but 
spinal fluid collected within the first 10 days 
of illness also may be cultured. Inocu- 
late the freshly-drawn specimen directly 
into several tubes of a suitable semisolid 
medium such as Fletcher’s. Multiple tubes of 
media should be used and small inocula, 1 to 
3 drops of blood to 5 ml. of medium, 
planted. As Fletcher’s medium remains 
stable for 3 to 4 months, the laboratory may 
supply it to the physician. It can be inocu- 
lated at the bedside, and then shipped by 
mail. Voided urine if diluted, may be 
cultured directly into semisolid medium. Five 
10-fold dilutions in buffered saline or a suit- 
able broth are first prepared. Since quantita- 
tive dilutions are not necessary, a single 
2.0 ml. syringe with a 20-gauge needle may 
be used to prepare these dilutions in the 
syringe. Draw up to 0.1 ml. of urine and 0.9 
ml. of diluent for the first dilution; expel all 
but 0.1 ml. of this dilution, planting one 
drop into 5 ml. of medium. For the second 
dilution, again draw 0.9 ml. of diluent into 
the syringe. Plant one drop of the mixture 
into 5 ml. of medium again expel all but 
0.1 ml. Repeat with the third, fourth, and 
fifth dilution. Otherwise, inoculate into such 
test animals as weanling hamsters or guinea 
pigs. 
Any pure cultures isolated should be shipped 
in semisolid media to a reference laboratory 
for confirmation by serologic tests, 
b. Specimens for serologic tests 
Microscopic or macroscopic agglutination 
tests are the most common serologic pro- 
cedures used for the diagnosis of leptospi- 
rosis. To detect a rise in titer, a specimen 
of blood should be taken at two different 
times: one during the first week or acute 
phase of illness, the other ten days or two 
weeks later. Maximum titers are usually 
reached by the third or fourth week. While 
serodiagnostic tests are of value in con- 
firming past or current leptospiral infection, 
paradoxical reactions may occur, and de- 
termination of the infecting serotype can be 
made only by isolation and serologic identi- 
fication of the leptospires. 
10. Meningitis. Any organism capable of invading 
human tissues can cause infection of the men- 
inges. As used here, the term “meningitis” refers 
to an inflammation of the meninges, brain, or 
spinal cord associated with increased cell counts 
in the spinal fluid. Characteristic of spinal fluid 
in bacterial meningitis is a predominance of 
polymorphonuclear cells and a decrease in 
dextrose content. 
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