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Alagdon, Edsel
Gomez, Paul Arvin
Laus, Abigail
BSMT 3-A
Tightly coiled, thin, flexible spirochetes
with both ends hooked
Motile
L. interrogans – all pathogenic strains
L. biflexa – saprophytic strains
Obligate aerobe
Culture media – 14% (vol/vol) rabbit
serum
• Fletcher’s semisolid
• Stuart liquid
• Ellinghausen-McCullough-Johnson-Harris (EMJH)
semisolid media
• Polysorbate 80 (Tween 80)-albumin
L. interrogans serovar
ictrohaemorrhagiae
• rats
L. interrogans serovar Pomona and
Hardjo
• cattles
Reduced Phagocytosis in the host
Cell-mediated sensitivity to
leptospiral antigen by the host
Small amounts of endotoxins
produced by some strains
Route – small breaks in the skin or intact mucosa
Incubation period – 10-12 days
Severe systemic disease (Weil’s disease) includes
renal failure, hepatic failure, and intravascular
disease, and may result in death.
Specimen – CSF or Blood/SDS-treated
blood (Acute phase)
• Urine can be culture after the first week of
illness
• Isolation
Direct inoculation of 1 or 2 drops of freshly drawn
blood or CSF into CM and incubate the media in
the dark at room temperature for 5 – 6 weeks at
28 ° C to 30 °C
Microscopic examination – Dark-field
microscopy (tightly-coiled, rapidly motile
spirochetes with hooked ends)
Urine (Specimen)
• It should be inoculated immediately
because the acidity of the urine might harm
the Leptospires.
• 1 or 2 drops of undiluted urine and 1:10
dilution of urine are added to 5 ml medium
• 200 ug/mL of 5-flurorouracil is added to
prevent contamination of other bacteria
Direct Observation
Insufficiently sensitive and extremely time
consuming
Presence of fibrin strands in blood
• In situ staining by immunofluorescence,
immunohistochemistry and sliver deposition
Used in detection of Leptospires in
veterinary histopathology
Replaced by PCR-based technology
Antigen Detection
• EIA, RIA and chemiluminescence
• Suffered from sensitivity problems and has
not gained widespread acceptance.
Itis a molecular diagnostic procedure
that has made possible the rapid and
sensitive detection of leptospiral DNA
in a variety of clinical samples.
The most frequent target sequences
are 16S and 23S rRNA genes.
IgM antibodies are detected within a
week after onset of disease.
IgG antibodies are detected a month
or more after onset of disease.
IgM dot-ELISA assay –
semiquantitative and it has 98%
sensitivity and 90.6% specificity.
MAT – Microsopic Agglutination test
• Gold standard for serological diagnosis.
• Positive result – agglution of live or killed Leptospires
assessed by dark field microscopy.
• It is most sensitive and specific serological test
available
MicroplateELISA – detect Leptospira IgM Abs
LEPTO dipstick and LEPTO Dri-Dot Test
Leptospires is susceptible in vitro to
antimicrobials streptomycin,
tetracycline, doxycycline, and the
macrolide antibmicrobics in vitro
Penicillin is considered beneficial and
alters the course of the disease if
treatment is initiated before the fourth
day of illness.
Doxycycline shorten the course of
illness in adults and reduce the
incidence of convalascent leptospiuria.
Measurement of hemoglobin, total
and differential white cell count,
platelet count
Renal Function tests
Serum bilirubin and serum
aminotransferase