Spirochetes

• Treponema
• Borrelia
• Leptospira
 Objectives
• To know primary, secondary, and tertiary
clinical manifestations of syphilis
• To know the diagnostic test to identify T.
pallidum in clinical laboratory
• To know the clinical picture of infection
caused by Leptospira organisms and to
identify them in the laboratory
• To know the clinical picture of infection
caused by Borrelia organisms and to identify
them in the laboratory
• Antimicrobial therapy
 Spirochete s

•Are spiral flexible organisms that move without flagella

•Multiply by transverse binary fission

Treponema : Many narrow regular coils

Borrelia : Few wide, irregular coils

Leptospira : Many very narrow coils with hooked ends

 Treponema pallidum
• Causative agent of syphilis : A STD
• Actively motile with an axial filament
• Are not stained with ordinary stains
• Are weakly refractile
Not seen with ordinary microscope
• Require dark-field microscopy to be
seen in fresh preparation
Growth requirements
• Microaerophilic
• Cannot be cultured on artificial media OR tissue culture
• Cultured by inoculation of rabbit testes
Treponema pallidum
 Treponema Pallidum

Antigen
• Outer-membrane protein (OMP) Ag
Antibodies : Two types
• Specific anti-treponemal Abs (IgM & IgG)
o Induced by OMP Ag
• Non-specific Abs (Reagin Abs-IgE)
o Stimulating Ag is not known
• Antibodies appear 2-3 weeks after
infection
 Syphilis

Source of Infection
• Patient with Primary or Secondary syphilis
Modes of Transmission
1. Venereal : Sexual contact
2. Non-venereal
A) Direct Contact
• With mucous membranes (kissing)
• Blood Transfusion
B) Mother to Child – Congenital syphilis
 Syphilis : stages
Primary Stage : Incubation period 2-4 weeks

Chancre Formation
Entry of organisms at site of contact
at site of contact After 3 weeks
(A flat dull ulcer)

Go to regional LN Chancre is painless &

(inflammed LN) Heals spontaneously
After 4-6 weeks

Chancre & LN
Bacteremia
Contain a large
no. of organisms
 Syphilis : stages

Secondary Syphilis :
• 6-12 weeks after chancre in untreated cases
• Generalized non-itchy, coppery-red rashes on
skin & MM especially on palms & soles
• Snail-track ulcers in oral cavity in 1/3rd of
patients
• The lesions contain many organisms
• Enlarged, painless, rubbery LN

Outcome in untreated cases

• 25% - Complete recovery
• 25% - Latent for life (Sero-positive)
• 50% - Tertiary syphilis
 SYPHILIS : stages

Tertiary Syphilis
• 3-10 years after primary lesion
• Chronic granuloma in skin, MM, bones or any
organ (gummas)
• Granuloma breakdown to form shallow ulcers
• Few treponemes in granuloma
• 10-20 years after primary lesion
o Neurosyphilis :
• Tabes dorsalis
• Cranial Nerve palsies
o CVS Changes : Aortic aneurysms, aortitis
 Syphilis : stages

Latent Syphilis
• Dormant for years
• Positive serology
Congenital Syphilis
• Trans-placental transmission from mother
with Pri. OR Sec. syphilis
• Fetal death OR Fetus borne with abnormalities
• Skin rashes, saddle nose, hepatitis, meningitis
Treatment
• Penicillin
 SYPHILIS : Lab Diagnosis

Specimens
• Primary Stage : Chancre & LN
• Secondary Stage : Skin rash & mucous patches

Microscopy
• Dark-field Microscopy
o To see motile treponemes
• Immunofluorescence

Serodiagnosis
 Syphilis : Lab Diagnosis
Serodiagnosis

Non-treponemal non-specific tests : To

detect non-specific Abs
• Extracts of normal mammalian tissues ( cardiolipin
from beef heart) react with anticardiolipin antibody
(Ab) in patient’s serum called Reagin
VDRL (Venereal Diseases research Lab) Test
• Slide agglutination test
• Rapid screening test
• Read microscopically
Rapid plasma reagin (RPR) Test
• Read with naked eyes
 SYPHILIS : Lab Diagnosis

• VDRL & RPR

• Are positive in majority of Pri syphilis
• Almost always positive in Sec syphilis
• Are sensitive but non-specific
• False positive in TB, Malaria,
Autoimmune diseases
• Have good prognostic value
• Titer is used to follow therapy
 Syphilis : Lab Diagnosis

Serodiagnosis
Specific tests
• Detect specific Abs
• FTA-Abs (Fluorescent treponemal antibody Absorption)
Test
• MHA-TP (MicroHaemAgglutination test for T. pallidum,
is an indirect hemagglutination test using T. pallidum
antigens absorbed to erthrocytes.

 A titre of non-specific Abs decreases with effective

treatment while specific Abs remain for life
 Syphilis : Treatment

• Penicillin
• Some patients with Sec syphilis,
experience
fever with chills & myalgias, a few hours
after penicillin.
• This response (Jarisch-Hexheimer) is due
to lysis of treponemes and release of
endotoxin-like substances.
 Nonvenereal Treponemes

 Bejel; caused by T. pallidum spp. Endemicum

• Common in middle east,

• causes granulomatous lesion of skin, bone and joint

 Yaws: Caused by T. pallidum spp. pertenue

• this disease occurs in tropical equatorial regions.

• causes granulomatous lesion of skin, bone and joint.

 Pinta: Caused by T. carateum

• Affect the skin with papules on the hands, feet and scalp. These
lesions heal slowly after treatment (unlike syphilis, yaws).
LEPTOSPIRA INTERROGANS

Leptospira : Many very narrow coils with hooked ends

• Causative agent of Leptospirosis : A

zoonotic disease
• Habitat : Infected domestic live-stock &
pets
• The organism settles in the kidney and
excreted in urine
 LEPTOSPIRA INTERROGANS

Mode of Infection
• Direct Contact with:
o Urine of infected animal
o Water & soil recently contaminated with infected
urine
• The organism enters body through:
o Skin lesions
o Conjunctival mucus membrane
o Ingestion
• High-Risk Groups
o Farmers
o Sewer workers
 Leptospirosis

Pathogenesis
• Incubation period : 2 day- 28 days
• Bacteremia : organisms multiply in liver,
spleen, kidney, meninges, conjunctiva
Clinical Features
• Influenza-like followed by hepatitis & meningitis
• Weil’s Disease (severe leptospirosis)
o Jaundice, hemorrhage, renal failure
Lab Diagnosis
• Dark field Microscopy of blood & CSF
• Sero-diagnosis
Leptospira sp
 LEPTOSPIROSIS

Treatment & Prevention

• Penicillin
• Animal immunization
• Proper treatment and disposal of
contaminated water
 Borrelia Recurrentis

Borrelia : Few wide, irregular coils

• Can be stained with Giemsa stain in blood film

• Can grow in media containing serum & tissue
extracts
• High frequency of antigenic variation in major
surface
protein and is responsible for :
o Organism can escape immune system
o Relapses of disease
• Disease : Relapsing Fever
Borrelia
 Relapsing Fever
Transmission :
• Tick-borne Relapsing Fever
o From infected rodents to human
• Louse-borne Relapsing Fever
o From infected human to human
Clinical Features
o Bacteremia : Infect various organs
o A non-pruritic rash at bite site
o Fever, rigors and headache for weeks to months
o Weeks to months later : Cardiac and neurological
symptoms like Bell’s Palsy, peripheral neuropathies
o Predominant arthritis
o One of the causes of PUO
 BORRELIA RECURRENTIS
Lab Diagnosis
• Giemsa staining of blood smear
• Detection of IgM OR rising titre of
IgG
• PCR

Treatment & Prevention

• Amoxycillin
• Louse, tick & rodent control
• Hygienic measures
 Borrelia Burgdorferi
• Cause lyme disease
• Fever, migratory skin rash
• Lyme disease rash called erythema migrans.
This rash often takes a bull's-eye
appearances and is observed in about 80% of
Lyme disease
• patients muscle and joint pain
• Evidence of meningeal irritaion
• In chronic cases, meningoencephalitis,
myocarditis and arthritis
• Transmitted to human by ticks
 Louse
Tick
Lab Diagnosis
• Detection of IgM OR rising titre of IgG
• PCR

Treatment & Prevention

• Doxycycline and beta-lactms
• Tick & rodent control
• Hygienic measures
 Features of Spirochetal Diseases

Organism Transmission Diagnosis

Microscopy Serology Disease

T.pallidum Sexual, Dark field of VDLR Syphilis
Transplacental chancre or sec. RPR,
Transfusion lesion FTA-ABS
MHA-TP
L.interrgans Ingestion of Not recommended Microagglutination Fever ,
contaminated (MAT) meningitis
water hepatitis
B. recurrentis lice Gemisa of blood None Relapsing
smear fever

B. burgdorferi ticks Not recommended EIA Lyme

disease
 Case #1
A farmer was admitted to hospital. He gave a
history of being unwell a week before admission
with fever and head ache. These symptoms
resolved, but the day before admission he
become pyrexial again and on examination was
found to be jaundiced and to have elevated blood
urea. Urine was collected and inoculated into
semisolid agar medium, which after 2 days
become turbid and was examined by dark ground
microscopy (see figure) Serum was also collected
for serology
 Questions

• What is your clinical suspicion of the

diagnosis?
• What further investigations would you
perform?
• How would you manage the patient?
 Case # 2
• A 24-year old Saudi student arrived from
New york, USA, at a local medical clinic
complaining of fever , fatigue and muscle
ache. It was mid September, and he had been
having symptoms for about 2 weeks. He
reported that he had removed numerous
insects, he assumed that some of them had
escaped detection for several days. On
examination the doctor noticed a circular
erythematous lesion on hid ankle, he said this
had followed an insect bite.
 Questions

• What is the insect and what relevance

does it have to his illness?
• What are the tests of choices for Lab
diagnosis of the disease?
• How should you treat this patient?
 Case # 3

A male patient presented at a

dermatology department with a history
of malaise and scaling, papular rash on
palms of his hands and an ulcer on his
hard palate. Further examination
revealed a generalized non-tender
lymphadenopathy. Dark ground
microscopy of scrapings of the mucous
ulcer revealed the organisms shown in
figure.
 Questions

• What was your diagnosis?

• How is the illness transmitted
• How should the patient treated?