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GROUP 6

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

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