Zoonoses

Leptospirosis

Prepared by:
Dr. Helen Tang Hooi Chia
Department of Microbiology (Faculty of Medicine)
 At the end of the lecture,
you should be able to:
❑ Describe the general properties
of the causative agent
❑ Describe the modes of
transmission and pathogenesis
❑ State the clinical manifestations
and complications
❑ Explain the laboratory diagnosis
❑ State the preventive measures
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Leptospira interrogans
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Causative Agent
Leptospira interrogans
❑ Gram-negative
❑ Aerobic spirochete

❑ Thin, flexible, elongated

❑ Tightly coiled spirals with hooked ends

❑ 5–15 μm long

❑ ________________
❑ Actively _________

❑ Many serovars based on cell surface antigens

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 Epidemiology
❑ Leptospirosis is worldwide in distribution
❑ 1.03 million cases worldwide annually with
58,900 deaths associated with the loss of 2.9
million disability-adjusted life years
❑ Most common in tropical areas and warmer
climates with lots of rainfall each year

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Mode of Transmission

Source of infection
❑ __________, dogs, cattle and pigs
❑ Asymptomatic animals can transmit
the infection via urine

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 Mode of Transmission
❑ ____________with ________ or other bodily fluid from infected animals

❑ ____________: water, moist soil and wet surfaces contaminated with infected animal urine
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Risk factors
❑ Occupational exposure:
Agricultural farmers, veterinarians, sewer workers, pet traders, military personnel, laboratory
workers
❑ Recreational activities:
Freshwater swimming, canoeing, kayaking

❑ Household exposure:
Pet dogs, domesticated livestock, rainwater catchment systems, infestation by infected rodents

❑ Low socioeconomic status:

Living in overcrowded urban areas with poor sanitation

❑ Others:
Walking barefoot through surface water, skin lesions, contact with
wild rodents, accidental laboratory exposure Manipal University College Malaysia 9
Pathogenesis
Portal of entry:
❑ Breaks in the skin

❑ Mucous membranes
❑ Ingestion (rare)

Two distinct phases of pathogenesis:

❑ First phase (____________ phase)

❑ Second phase (__________ phase)

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First phase (septicemic phase)
❑ L. interrogans spill over to the bloodstream
❑ Disseminate hematogenously to various organs
(brain, lung, heart, liver and kidney)

❑ _______________: Spirochetes can be found in the

walls of capillaries, medium and large-sized vessels
Pathogenesis
❑ Penetration and invasion of tissues is due to active
motility and release of hyaluronidase
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Second phase (immune phase)

❑ Develops when the IgM antibody titer
rises
❑ Clearance of L. interrogans occurs
when antibodies develop
❑ Antigen-antibody complexes are
deposited in various organs

❑ ________________: Bacteria become

adherent to the proximal renal tubular
brush border and are excreted in
urine

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 Incubation period: around 10 days (1-30 days)
Clinical Manifestations Clinical manifestations: produces two distinct clinical syndromes
& Complications ❑ Mild anicteric febrile illness
❑ Weil’s disease (Hepato-renal-hemorrhagic syndrome)

1-30 days

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 Clinical Manifestations & Complications

14 Mild anicteric febrile illness

Weil’s disease
(Hepato-renal-hemorrhagic syndrome)

❑ 90% ❑ 10%
❑ ____________ ❑ Typical biphasic course may not be present
❑ Flu-like illness with fever, ❑ It often progresses rapidly with a case–
chills, headache, nausea, fatality rate of 5 to 15%
vomiting, abdominal pain, ❑ Icterus (jaundice), high grade fever,
myalgia, and conjunctival hemorrhagic manifestations and impaired
suffusion renal
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Clinical presentation of leptospirosis

a Subconjunctival hemorrhages and icterus in a 37-year-old

man who kept pet rats presented with sudden onset of fever,
myalgia, and severe headache

b Severe pulmonary hemorrhage in a 50-year-old man who

had recently returned from vacation in Malaysia where he
had waded through mangrove forests.

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 Laboratory Diagnosis

Diagnosis
❑ Microscopic
❑ Culture isolation
❑ Animal inoculation
❑ Serology
❑ Molecular methods
❑ Non-specific findings

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 Specimens' collection
First phase Second phase
❑ CSF
❑ __________ ❑ _________

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Microscopic

Cannot be seen under light microscope

Wet films: Observed under dark field or phase contrast microscope
Staining: Sliver impregnation stains (Fontana stain and modified Steiner Technique)

Interpretation
• Reveals spirally coiled bacilli
• Hooked ends like umbrella handle
• Highly motile; exhibit spinning and translational movement

Disadvantages
• Less sensitive
• Requires technical expertise
• Serum proteins and fibrin strands in blood may resemble leptospires
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Culture Isolation
❑ Laborious
❑ Technically demanding
❑ Time-consuming
Culture condition
• Leptospira is an obligate aerobe and slow growing
• Incubated at 30oC for 4–6 weeks Schematic diagram (viewed under dark-
field microscope)
• Examined under the dark field microscope
Culture media
As Leptospira is highly fastidious, requires enriched media
(1) EMJH liquid/semisolid medium (Ellinghausen, McCullough, Johnson, Harris)
(2) Korthof’s medium
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 Dinger’s ring
The opaque ring just below the
surface of the semi-solid EMJH
media in the tube with positive
culture

Growth of leptospires in EMJH liquid (a) growth &control

and EMJH semisolid medium (b) control &growth.
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Serology
Genus-specific tests __________ specific tests
• Uses broadly reactive • Microscopic
genus-specific antigen agglutination test
prepared from non- (MAT)
pathogenic strain
MAT serves as the gold

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• Cannot detect the standard reference
infecting serovar method

• Macroscopic slide
agglutination test, latex
agglutination test, ELISA,
ICT
Laboratory Diagnosis
Animal inoculation
Samples are inoculated into hamsters and young guinea pigs

Molecular method
❑ PCR
• Not serovar-specific
Non-specific finding
❑ Altered renal function
Elevated levels of blood urea nitrogen and serum
creatinine

❑ Altered liver function

Elevated bilirubin and liver enzymes in serum Manipal University College Malaysia 23
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Budihal SV, Perwez K. Leptospirosis diagnosis: competancy of various laboratory tests.

J Clin Diagn Res. 2014 Jan;8(1):199-202. doi: 10.7860/JCDR/2014/6593.3950
 Based on awareness of leptospirosis epidemiology and
transmission mechanisms
❑ Chemoprophylaxis with doxycycline is recommended for
anticipated short-term exposures
❑ General sanitation approaches (proper waste disposal)

❑ Rodent control
❑ Do not swim in water that might be contaminated
with animal urine
❑ Health education

Vaccine
❑ Whole-cell vaccines (mono- or polyvalent)
containing specific serovars of Leptospira

Preventive ❑ SPIROLEPT: For subcutaneous injection

Measures ❑ The efficacy rate is around 60–100% 25
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