1. DON XAVIER N.D
SPIROCHAETES

2. MORPHOLOGY
Body have Central cytoplamic cylinder bound by cytoplasmic membrane and cell wall having Gram –ve properties.
Presence of internal Filaments called ENDOFLAGELLA.
Show three Types of motility
Cork screw like
Flexion & extension
Translatory motion

3. TREPONEMA PALLIDUM FEATURES
Spirochaete with filamentous helices with tapering ends and extremely thin (0.2um).
Cannot be seen by wet film or Aniline dyes and only by Dark field microscopy.
In Silver impregnation method it reduce silver nitrate to metallic silver and get deposited on surface.
Cannot be grown in media, but can be grown in Rabbit testicle and other Epithelial cells.

4. TREPONEMA PALLIDUM PATHOGENSIS
Two major types SPECIFIC and NON SPECIFIC ANTIGEN types.
SPECIFIC ANTIGENS
Group specific Antigens--- All Treponemas have group specific antigens, antibody to which is found in serum of Syphlitic patients.
Species specific antigens---- Appears to be Polysacchride in nature.
NON SPECIFIC ANTIGENS (Reagin)
It occurs in blood of Syphilitic patient that react with lipid hapten antigen , DIPHOSPHATIDYL GLYCEROL called Cardiolipin.

5. Virulence factors are less discovered.
Outer membrane proteins
Hyaluronidase
Fibronectin – attachment of organism to host cell is by absorption of fibronectin to organism.

6. TREPONEMA PALLIDUM LABORATORY DIAGNOSIS DIRECT DEMONSTRATION
Dark field microscopy (DGI)
Stained Preparation – Silver impregnation method ( FONTANA’S METHOD)
Tissue Biopsy – Stained by Levaditis stain or Warthin Starry stain
PCR

7. 0.05 ml decomplemented serum
SEROLOGICAL TESTS
Non Treponemal Tests – Cardiolipin Antigen (Alcoholic extract of bovine heart muscle with lecithin and cholesterol) + Reagin ( from patient’s serum) –FLOCCULATION.
Before test patients serum is inactivated by heating at 56’c to destroy complements.
VDRL TEST-
0.05 ml decomplemented serum
Add cardiolipin antigen drop by drop till 1ml slide is rotated for 180 revolutions per minute.
antigen antibody reaction.

8. Same as RPR but give more stability in hot climatic conditions
RPR TEST
Same as VDRL test where finely divided charcoal and Choline are added to stabilize VDRL antigen.
TRUST TEST
Same as RPR but give more stability in hot climatic conditions
COMPLEMENT FIXATION TEST
0.05 ml decomplemented serum
Add cardiolipin antigen
Serum from guniea pig (complement) is added
Incbate for 1 hr in 37’c
If reagin is present it fixes the complement
sRBC are added –it consists of mixture of sheep RBC suspension
And anti-sheep erthrocyte serum antibody(amboceptor)
Incubate for 1 hr in 37’c

10. BORRELIA

11. BORRELIA
FEATURES
Largest of spirochaetes , so stained easily as gram negative.
Have 5-8 irregular spirals at interval of 2um with pointed ends.
Diseases commonly associaed with them are :
Relapsing fever– Epidemic louse borne –B. recurrentis
Endemic tick borne – B.duttoni
Lyme Disease - Tick borne – B.burgdoferi
Can be grown in serum,blood,or tissue enriched liquid medium
Cultivated in chorio-allantoic membrane of chick, Peritoneal cavity of rat or mice.

12. BORRELIA RF PATHOGENSIS DISSEMINATION
After arthropod bite, Borrelia spread by blood stream to mutiple organs.
RECCURENCE
Occurs due to antigenic variation and modulation, an immunological phenomenon.

13. BORRELIA RF LABORATORY DIAGNOSIS Smears Blood Cultures Serology
Animal Inoculation

14. BORRELIA LD FEATURES Flexible, helical, gram negative
Caused B.burgdoferi
Tick borne infection

15. BORRELIA LD PATHOGENSIS
B.burgdoferi have toxic lipopolysacchride in the cell wall and peptidoglycan have inflammatory properties.
After bite one or More skin lesions develpo at site of skin infection
Organism migrate out by lymph channel and reaches
Blood stream
Cause late manifestations as cardiologic and neurologic conditions , increase in serum IgM levels and immune complexes.
Causes Arthritis and deposition of immune complexes in synovial Tissue.

17. LEPTOSPIRACEAE

18. LEPTOSPIRA
FEATURES
Spiral organisms with numerous set coils and hooked ends
Narrow diameter (0.1um)
Actively motile in liquid media,by spinning motion.
Do not Stain by Gram’s stain.
Observed by DGI, Silver stain.

19. LEPTOSPIRA PATHOGENSIS L.interrogans cause of Zoonotic disease
TRANSMISSION
By direct or indirect contact by urine and Faeces of carrier animals.
PORTAL OF ENTRY
Enter body by Cuts or Abrasions on skin, or by intact membrane of mouth, nose or eye.
DISSEMINATION
After incubation of 1-7 days it enter the blood stream and disseminates haematogenously to variety of organs and tissues to cause jaundice, increased transaminases, DIC, Renal failure.

20. LEPTOSPIRA LABORATORY DIAGNOSIS
Blood and Urine are taken as specimens with serum, CSF
ISOLATION
very difficult and cultured in Fletcher’s Medium by culturing
centrifuged urine. Incubated for two to three weeks at 30’c.
DGI , MAT
CFT
ELISA – to detect IgM or IgG seperately.
CONFIRMATORY TESTS --- Serogroup specific antigens
Live or Formalin killed suspension of formalin killed suspension of leptospires are prepared , a series of patients serum dilution is tested against the antigen.