SPIROCHAETES

Wasswa Fredrickson B
2021/PhD/047/PS
 Definition
• Long helical, corkscrew-shaped motile
bacteria with tight or loose spiral depending
on the spp

• Gram negative measure 0.1 to 3 µm by 5 to

120 µm.

• Replication is slow (doubling time, 30 hours)

 Taxonomy
• The order spirochaetales has two families
• The family spirochaetaceae- which consist of free living large spiral
organisms and
• The family Treponemataceae has three main genera: causing disease
in man namely
• i) Treponema
• Ii) Leptospira and
• Iii) Borrelia
• The genus Treponema has two spp that cause human disease
• i) Treponema pallidum with 3 subspp
_ T.P Pallidum → Syphilis
– T. p. endemicum → Endemic syphilis (Bejel)
– T.p. pertenue → Yaws
ii) Treponema carateum → Pinta
 Taxonomy
The genus Borrelia has the following medical important spp
• i) Borrelia burgdorferi → Lyme disease
(borreliosis)
• Ii) Borrelia recurrentis → Epidemic relapsing fever
(louse borne relapsing fever)
• Iii) Other Borrelia species → Endemic relapsing fever
(Tick-borne relapsing fever)
• Iv) B,vincenti → Together with fusifrom bacteria causes ulcerative
gingivo stomatitis (vincent angina)
The genus Leptospira interrogans
• Two spp
• i) L. interogans - pathogenic
• Ii) L. biflexa - free living non pathogenic
• → Leptospirosis (Weil’s Disease)
 Structure
• The organisms are covered by the outer sheath
(glycosaminoglycan)

• The outer membrane contains peptodiglycan and maintain the

structural integrity of the organism

• The endoflagela (axial filaments) are flagella like organelles in

the periplasmic space encased by outer membrane.

• The endoflagella begins at each end winds around it extending

to and overlap at the midpoint

• Inside the endofloagella is the inner membrane (cytoplasmic

membrane) that provide osmotic stability and covers the
protoplasmic cylinder

• They reproduce by tranverse fission.

Structure
 Treponema
Morphology
•Slender spirals measuring 0.2 µm in width and 5-15 µm in
length
•They have tight spirals i.e are regularly spaced at a distance of
1 µm
•Have pointed and tapering ends,
•3-4 endoflagella, actively motile, show rotatory corkscrew like
movement
•The spiral are thin require immunofluoresence or dark field
illumination, Stained by silver impregnation
•Microaerophilic surviving at 1-4% Oxygen
•In proper suspending fluid and presence of reducing agents
survive for 3-6 days at 25oC or in blood at 4oC for 24 hrs
•They stain with difficulty except with Giemsa's stain or silver
impregnation.
Treponema: Regular spiral coils 1 µm
apart
 Treponema
Culture and growth characteristics
•T pallidum has never been cultured continuously in artificial
culture media
• T. pallidum is usually cultured in the testes of rabbits,
• Non pathogenic /saprophytic Treponema can be cultured
anaerobically invitro doubling time is 30 hrs. The saprophytic
Reiter strain grows on a defined medium of 11 amino acids,
vitamins, salts, minerals, and serum albumin.
• T pallidum is a microaerophilic organism; it survives best in 1–
4% oxygen. In proper suspending fluids and in the presence of
reducing substances, T pallidum may remain motile for 3–6
days at 25 °C.
• In whole blood or plasma stored at 4 °C, organisms remain
viable for at least 24 hours, which is of potential importance in
 Scientific Classification
Scientific classification
Domain: Bacteria
Phylum: Spirochaetes
Order: Spirochaetales
Family: Spirochaetaceae

Genus: Treponema
Species: T. pallidum

Binomial name
Treponema pallidum
Schaudinn & Hoffmann, 1905
 Treponema
Reactions to Physical and Chemical Agents
• Drying kills the spirochete rapidly, as does elevation of the temperature to
42 °C.

• Treponemes are rapidly immobilized and killed by trivalent arsenical,

mercury, and bismuth (contained in drugs of historical interest in the
treatment of syphilis.

• Penicillin is treponemicidal in minute concentrations, but the rate of killing is

slow, presumably because of the metabolic inactivity and slow multiplication
rate of T pallidum (estimated division time is 30 hours).

• Resistance to penicillin has not been demonstrated in syphilis.

 Treponema
• Virulence Factors
• Several gene products associated with virulent strains,
– Their roles in pathogenesis are unknown.

• The outer membrane proteins are associated with adherence to the

surface of host cells,

• Virulent spirochetes produce hyaluronidase, which may facilitate

perivascular infiltration.

• Virulent spirochetes are also coated with host cell fibronectin, which
can protect against phagocytosis.
 Treponema
• 1. T.p. pallidum – Syphilis
Pathogenesis and pathology
• Syphilis is an acute and chronic infectious disease
caused by Treponema pallidum.
Routes of infection:
• Mucosal membranes of the vagina or urethra,
sometimes scar wound or scratches.

• Across the placenta to the developing fetus

• T. pallidum tends to invade the interstitial spaces of

tissue at the site of infection and to move rapidly to
other locations (active "cork-screw" motility)
 Treponema
Clinical presentation
1.Primary Syphilis:
- Following penetration of the skin or mucous membranes, a
characteristic, painless hard chancre develops at the site of entry
within 3 weeks (10-60 days after exposure)at the primary site of
initial replication: penis, labia, cervix, anorectal region, the mouth.
- The chancre is hard, slightly elevated, round, ulcerous and highly
contagious filled with Treponemes.
- Simultaneously the organism enters the lymphatics and becomes
disseminated.
- The chancre heals without treatment in a few weeks(4-6 weeks)
due to local immunity, even without treatment, may leave a scar but
by that time the organism has already disseminated.
 Treponema
Primary Syphilis:
• The lymph nodes also become inflamed.
• Histology endarteritis (inflammation of the inner
wall of an artery) and periarteritis
• Inflammation of membranous sac surrounding the
heart with polymorphonuclear leukocytes and
macrophages.
• 1:3 of people exposed becomes infected.
• Unless syphilis is treated early in this stage, it will
progress into secondary syphilis.
Primary syphilis
 Treponema
Secondary Syphilis:
•4-8 weeks after the chancre heals,
•Typically there are lesions (filled with treponemes) throughout the
body including the skin, mucous membranes, organs, and eyes.
•Lesions appear as pale red rash appears usually on the palms or soles
of feet, or over the entire body.
•Accompanied by fever, sore throat, headaches, joint pains, poor
appetite, weight loss, and hair loss.
•Sores form around the genitals or anus that secretes extremely
infectious fluids.
•These symptoms usually last 3-6 months, but may not last and then re-
appear at any time.
•Without treatment, syphilis will continue to progress throughout the
body.
•This also heals without treatment and the patient may either
spontaneously get well or develop a latent infection
Secondary syphilis
 Treponema
The Latency Stage:
•No obvious symptoms but specific anti-treponemal
antibodies are found.
•T. pallidium is lodging itself into the host's tissue.
•This stage may last from few months , 3-10 years.
•Generally individual is not infectious
•Always passes the disease to the fetus.
•Approximately 1/3 to ½ may eventually progress to the next
stage.
•50-70% will not progress to tertiary syphilis.
 Treponema
Tertiary Syphilis:
•This stage is characterized by granulomatous lesions, called
Gummas of which can occur on the skin, internal organs, CNS,
bones, eyes, and cardiovascular system. the heart, brain and liver,
can be fatal.
• They cause the body’s hyperimmune reaction to remaining
spirochetes.
• When lesions develop in the CNS it is called neurosyphilis and it
can lead to paralysis.
• In the eyes it can lead to blindness, and in the heart to aortic
damage or aneurisms.
•Gumma lesions whose pathogenesis is uncertain frequently lead
to destruction of soft tissue or bone.
•Occur for an extended period from as few as 2 to over 40 years
from onset of infection.
Tertiary syphilis
 Treponema
Congenital Syphilis:
•Occurs when the treponemes cross the placenta during the fifth
month to infect the unborn fetus (occurs usually when mom is in
the latent stage).
•This can result in damage to mental development or other
neurological symptoms or the child may be born with generalized
syphilis.
•In a pregnant woman who has a primary or secondary stage of
the disease, this usually results in stillbirth
•Multi-organ deformities or latent infections.
•Most infected infants are born without clinical evidence of
disease,
– May develop rhinitis followed by a widespread papular rash.
– Late bony destruction and cardiovascular syphilis
 Treponema
Diagnosis
•Specimen: Secretions (in ulcers), serum, cerebral spinal fluid
1. Direct examination (Microscopy)
•Dark field illumination: motile spirochetes
•Direct Immunofluorescence antibody staining (FA)
– Stained using flourescein-labelled antitreponeme
serum : fluorescent spirochetes
2. Culture: Efforts to culture T. pallidium have been generally
unsuccessful.
3. Serology
•Non-specific; non-treponemal tests
•Specific treponemal tests.
 Treponema
Non-treponemal tests:
•Detect IgG and IgM antibodies (reagin antibodies) developed against lipids from
damaged cells during the early stage of the disease.
•The antigen used for the nontreponemal tests is cardiolipin, derived from beef heart.
– Venereal Disease Research Laboratory (VDRL)
– Rapid plasma reagin (RPR) test.
– Coagulation of cardiolipin antigen by the patient's serum.
– Both tests are rapid,
– VDRL test can be used to test cerebrospinal fluid from patients with suspected neurosyphilis
– Become negative after successful treatment
Treponemal tests
– Specific antibody tests used to confirm positive reactions with the VDRL or RPR tests.
– Can be positive before the nontreponemal tests in early syphilis.
– Remain positive when the nonspecific tests revert to negative in some patients who have late
syphilis.
•→ Fluorescent Treponemal Antibody Absorption (FTA-ABS) test
•→Treponemal pallidum Hemagglultination Test (TPPA, TPHA)
• Treponema pallidum –Spp. that responsible for
syphilis.
• From family of Spirochete
• Pathogenic treponemes:
• -T. pallidum
• -T. carateum
• -T. pertenue
• -T. cuniculi
• -T. endemicum
 Antibodies
• May be directed against T. pallidum or
protein antigen group common to
pathogenic treponemes
• IgM in early or untreated latent syphilis
• Rapidly followed by IgG w/c becomes
predominant during secondary stage
 Non Treponemal VS. Treponemal
• Non – treponemal indirect method • React in Treponema pallidum strains.
• Detect antibodies specific to T.
of blood test for diagnosis.
pallidum.
• Tests detect biomarkers that are • Although these tests are typically
more specific than nontreponemal
released during cellular damage.
assays, they will remain positive.
• Nonspecific antibodies that react .
to cardiolipin.
• Measure immunoglobulin.
• Ideally used for qualitative syphilis
screening.
• CDC approved test include VDRL,
RPR.
 Non-Treponemal test Treponemal test
Flocculation Tests Treponema
Rapid Plasma Reagin Hemaglutination
(RPR) Absorption(TPHA)
 Venereal Disease Research Fluorescent treponemal
Laboratory(VDRL) antibody absorption (FTA-
ABS)
Cardiolipin Treponemal
 Serological Test for SYPHILIS

• VDRL (Venereal Disease Research Laboratory)

• ELISA (Enzyme-Linked Immunosorbent Assay)

• RPR (Rapid Plasma Reagin)

• FTA-ABS (Fluorescent Treponemal Antibody

Absorption Test)
• TPI (Teponema pallidum Immobilization Test)

• TPPA (Treponema pallidum Particle

Agglutination Assay)
• MHA-TP (Microhemagglutination Assay)

• DARK FIELD MICROSCOPE

 VDRL
• Screening test for syphilis, routine part of prenatal care during
pregnancy .

• Several states also require screening for syphilis prior to obtaining

a marriage license.

• Measures Antibodies.

• Specimen for the test is SERUM

• Screening test is most likely to be positive in the secondary and

latent stages of syphilis

• Test is not accurate in Early and Tertiary stage.

• Positive (Reactive)

• Negative result it means (Non-Reactive)

• Confirm the results with an FTA-ABS test, which

is a more specific syphilis test.
• Body does not always produce antibodies.
 ELISA
• Reaction is performed on polystyrene, the reaction is
between the Antigen and Antibody
• Polystyrene carriers which have absorbed an
ultrasonicated preparation of T. Pallidum.
• Positive result the carriers (Red-brown-violet)

• Negative (White) does not change

• Check antibodies for syphilis

 RPR
• RPR test is used in screening of infection, athough treatment can
begin to spread of the disease by an infected to unaware
person.
• Similar test to the VDRL.

• Detection present of non-specific antibody.

• Second stage or middle stage is one of the most accurate stage

to perform RPR
• FTA-ABS is confirmatory test.
• Large clamp (Positive)
• NO clumps (Negative)
•
FTA-ABS
Using antibodies specific T.palllidum

• Such tests are more specific than non-treponemal testing

• Positive earlier and remains positive longer than VDRL

• Should always be followed to confirm a positive RPR and/or VDRL test

for syphilis

• Highly sensitive but time consuming

• w/fluorescence: REACTIVE; w/o fluorescence: NONREACTIVE

 TPI
• Mixing of live Reiter treponemes from a
testicular chancre of rabbit and patient sera
• Immobilization of treponemes = REACTIVE
• Treponemes remain motile = NONREACTIVE
• Observed under dark-field microscope
• Standard test detecting NEUROSYPHILIS.
 TPPA
• Indirect agglutination assay used for detection and
titration of antibodies.
• Gelatin particles are sensitized with T. pallidum antigen.

• A negative test shows no clumping of gelatin particles.

• A positive test no clumping of gelatin.

• TPPA has a sensitivity of 85% to 100%, and

a specificity of 98% to 100%. For 1st stage of syphilis.
• Confirmatory test
Treponema pallidum
Particle Agglutination
 TPHA & MHA-TP
• Uses tanned sheep RBCs coated with
antigens from Nichols strain of TP
 Dark Field Microscope
• Direct detection of the organism and indirect evidence of
infection.

• Molecular assays to detect T pallidum DNA, and histopathologic

examination of biopsies of skin or mucous membranes.

• Allows visualization of live treponemes.

• Detection on infection before the serological test.

• corkscrew appearance (+)

• + result may indicate type of any stage Syphilis

 Treponema

Treatment, Prevention and Control

•Penicillin is the drug of choice since in 1943
– Benzathine penicillin or penicillin G
•Tetracycline, erythromycin, and chloramphenicol
•Penicillin or chloramphenicol in neurosyphilis.
Epidemiology
•Prevalence in Tanzania 4-7% in pregnant women
•Associated with HIV transmission
Prevention:
•Health Education to include abstinence, condoms use
 Treponema

• Other diseases related to syphilis

• Caused by organisms related to T pallidum,
• Give positive treponemal and non treponemal reactions
• Spread by direct contact only not (sexually)

1 Bejel
• Caused by T pallidum subspecies endemicum
• Endemic in Africa, Middle east, SE Asia
• Highly infectious skin lesions; late visceral complications
• Rx Penicillin
 Treponema

2 Yaws
•Caused by T pallidum subspecies pertenue
•Endemic in children in tropical countries
•Ulcerative lesion on the arms and legs
•Scar formation and bone lesions are common
•A variant of syphilis??
•Rx Penicillin
3 Pinta
•Caused by T carrateum
•Endemic in all age groups in Mexico, Central and S America
•Non ulcerative papule on the skin followed by hyper-pigmented
lesions
•Rx Penicillin
THANK YOU!