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  1. 1. Spirochetes Gram -ve
  2. 3. <ul><li>Elongated ,motile,flexible,twisted spirally along the long axisendoflagella-polar,wound around helical potoplastic cylinder—between outer membrane and cell wall. </li></ul>
  3. 5. Treponema <ul><li>Short,slender,fine spirals,pointed/round ends </li></ul><ul><li>Both commensals and pathogens </li></ul><ul><li>Venereal syphilis-T.pallidum </li></ul><ul><li>Endemic syphilis-T.pallidum/T.endemicum </li></ul><ul><li>Yaws-T.pertenue </li></ul><ul><li>Pinta-T.carateum </li></ul>
  4. 6. <ul><li>Treponma pallidum </li></ul><ul><li>Pallidum-pale staining </li></ul><ul><li>Morpho-thin ,delicate,ten regular spirals-sharp and angular(indian ink),motile(dark ground/phase contr)-sec. spirals appear & dis appear-but primary persists </li></ul><ul><li>Silver impregnation method,fontana’s method,Leviditi’s method(tissue sections) </li></ul><ul><li>Culture-do not grow in artificial media </li></ul>
  5. 8. <ul><li>Virulent strains-maintained by serial testicular passage in rabbits for decades </li></ul><ul><li>In culture-isolates are mainly non-pathogenic-but similar morph. And antigenic property </li></ul><ul><li>Best known is REITER STRAIN –Tr.phagedenis </li></ul><ul><li>Imp for specific tr. Test for identification of syphilis </li></ul>
  6. 9. <ul><li>Resistance-delicate,fomites hav no influence in transfer </li></ul><ul><li>Fever therapy -killed in 1 to 3 days at 0-4C,so transfusion syphilis can be prevented by storing blood for 4 days </li></ul><ul><li>Natural infection occurs only in humans </li></ul>
  7. 11. <ul><li>Antigenic structure-treponemal infection—three types </li></ul><ul><li>Reagin-STS/Nonspecific tests-antigen is hapten/cardiolipin(diphosphatidyl glycerol) from beef heart </li></ul><ul><li>Group antigen-in pathogenic and non pathogenic </li></ul><ul><li>Species specific-PS antigen-only positive in patients infected with pathogenic treponems </li></ul>
  8. 13. <ul><li>Primary- chancre-at the site of entry </li></ul><ul><li>Painless, realtively avascular,superficial ,indurated,ulcerated lesion. </li></ul><ul><li>Hard chancre-/Hunterian chancre—covered by exudate rich in spirochetes </li></ul><ul><li>Regional lymph nodes are swollen,discrete,rubbery and non tender </li></ul><ul><li>Chancre heals by 10-40 days even without t/t </li></ul>
  9. 14. <ul><li>Secondary -1-3 months after first </li></ul><ul><li>Due to widespread multiplication and dissemination in blood. </li></ul><ul><li>Skin rashes,mucus patches,condylomata </li></ul><ul><li>Spirochetes are abundent in the lesions,patient is most infective </li></ul><ul><li>There may be ophthalmic,osseous,meningeal involv. </li></ul><ul><li>Spontaneous healing over many years </li></ul>
  10. 15. <ul><li>Latent syphilis-period of quiescence after sec.syphilis </li></ul><ul><li>Natural cure may follow----but may devlp tertiary syphilis </li></ul><ul><li>Teritiary syphilis-cardiovascular lesions including aneurysms,chronic granulomata/gummata,meningovascular manifestations </li></ul><ul><li>Lesion contains few spirochetes,it is manifestation of delayed hypersensitivity </li></ul><ul><li>Also-tabes dorsalis,paralysis-several decades after -late teritiary/quartenary </li></ul>
  11. 16. Complications <ul><li>Neurosyphilis </li></ul><ul><li>Cardiovascular syphilis </li></ul><ul><li>Gummatous syphilis </li></ul>
  12. 17. <ul><li>Occupationally acquired-primary chancre in fingers </li></ul><ul><li>Blood transfusion-absent </li></ul><ul><li>Congenital- lesions develop only fourth month of gestation-when fetal immune competence starts appearing-pathogenesis requires immune response from fetus </li></ul><ul><li>Can b prevented if treated before 4 th month </li></ul><ul><li>If untreated the obstetric history will b-one abortion,still birth,live birth with syphilis,and finally healthy infants </li></ul>
  13. 18. <ul><li>Lab dignosis </li></ul><ul><li>Demonstration of spirochets </li></ul><ul><li>Antibodies in serum /csf </li></ul><ul><li>Microscopy –for primary,secondary,congenital cases---dark ground microscope </li></ul><ul><li>Identified by slender spiral structure and slow movement-but 10*4 per ml is needed for the test to be +ve </li></ul><ul><li>Direct fluorescent antibody test for TP is the best (DFP-TP) </li></ul>
  14. 19. Serological tests <ul><li>Reagin Antibody test </li></ul><ul><li>Group specific treponemal test </li></ul><ul><li>Specific Trponema pallidum test </li></ul>
  15. 20. STS/ Reagin antibody test Wassermann complement fixation test Kahn flocculation test VDRL Rapid Plasma Reagin
  16. 21. <ul><li>VDRL-inactivated serum(serum heated at 56C for 30 min)-is mixed with cardiolipin on a special slide—rotated for 4min---positive-visible clumps-by serial dilution titre can be known </li></ul><ul><li>VDRL Can be tested for CSF, but not plasma </li></ul><ul><li>Rapid plasma Reagin-VDRL antigen with fine carbon particles-more clear result-can be done with unheated serum /plasma—but not with CSF </li></ul><ul><li>Reagin antibody becomes detectable 7-10 days after appearance of primary chancre </li></ul><ul><li>/ 3-5 weeks after acquiring infection </li></ul>
  17. 22. <ul><li>Biological false positive in STS </li></ul><ul><li>Acute infections, injuries, inflammations-recheck after it </li></ul><ul><li>SLE and other collagen d/s </li></ul><ul><li>Leprosy </li></ul><ul><li>Malaria </li></ul><ul><li>Relapsing fever </li></ul><ul><li>IMN </li></ul><ul><li>Hepatitis </li></ul><ul><li>Tropical eosinophilia </li></ul>
  18. 23. <ul><li>Group Specific Treponemal tests </li></ul><ul><li>Reiter protein Complement Fixation-using a lipopolysaccharide-protein complex antigen derived from the treponeme </li></ul><ul><li>Free of BFP </li></ul><ul><li>Not in general use </li></ul>
  19. 24. Specific T Pallidum Test TP Immobilisation test Fluorescent T antibody test TP haemagglutination assay FTA-ABS-absorption
  20. 25. <ul><li>Specific T P test-use virulent Nichol’s strain of TP, maintained by serial inoculation in rabbit testis-renderd nonmotile if antibodies present </li></ul><ul><li>FTA-prepared antigen slides from nichol’s strain-test serum added-studied the fluorscence </li></ul><ul><li>FTA-ABS-test serum is preabsorbed with a sonicate of Reiter treponemes(sorbent) to eliminate group specific reactions </li></ul><ul><li>TPHA_tanned RBC senitised with a sonicated extract of TP as antigen </li></ul><ul><li>T/t-penicillin/doxacycline </li></ul>
  21. 26. Non veneral Trepanomatoses <ul><li>Endemic syphilis- CF similar to sec.syphilis—gummatous lesion—complications rare </li></ul><ul><li>Yaws -primary-extra genital papule-enlarges & breaks to form ulcerating granuloma-sec.and tert,.—compli.—rare-destructive gummatous bony lesions are common </li></ul><ul><li>Pinta-primary-extra genital papule-develops into psoriaform patch—sec-hyper/hypo pigmentation in skin </li></ul>
  22. 27. <ul><li>Non veneral treponema— </li></ul><ul><li>T.denticole </li></ul>
  23. 28. Borrelia <ul><li>Large ,motile,refractile spirochetes with irregular ,wide open coils </li></ul><ul><li>Those causing Relapsing fever </li></ul><ul><li>B vincenti -fusospirochetosis </li></ul><ul><li>B burgdorferi –Lyme disease </li></ul>
  24. 29. Relapsing fever <ul><li>Louse borne-B.recurrentis-vector-pediculus humanus corporis </li></ul><ul><li>Tick borne-accidental human infection </li></ul><ul><li>Antigenic prop-readily undergoes antigenic variations in vivo-that’s y relapses occurs-by DNA rearrangements in linear plasmids </li></ul><ul><li>Ultimate recovery by developiong immunty against all </li></ul><ul><li>Detected by lashing movemenmt </li></ul><ul><li>RF- false positive for syphilis </li></ul><ul><li>Agglutinins for proteus OXK are sometimes seen in high titres in louse born </li></ul><ul><li>T/t-tetracycline,penicillin,chloram,erythro </li></ul>
  25. 30. Borrelia vincenti <ul><li>Normal mouth commensal </li></ul><ul><li>Under malnutrion- </li></ul><ul><li>Ulcerative gingivostomatitis/oropharyngitis/Vincent’s angina </li></ul><ul><li>In these cases,it is associated with fusiform bacilli(Fusobacterium fusiforme)-this symbiotic infection is known as Fusospirochetosis </li></ul><ul><li>T/t-penicillin,metronidazole </li></ul>
  26. 31. Lyme Disease <ul><li>By B.burgdorferi </li></ul><ul><li>Three stages </li></ul><ul><li>Erythema migrans-expanding annular skin lesions </li></ul><ul><li>Disseminated infections-fever,headache,myalgia,arthralgia,lymphadenopathy </li></ul><ul><li>Pesistant infection-c/c arthritis,polyneuropathy,encephalopathy and acrodermatitis </li></ul><ul><li>Vector-ixodes dammini – tick </li></ul><ul><li>t/t doxycycline </li></ul>
  27. 32. Leptospira <ul><li>Actively motile, delicate,large no. of closely wound spirals </li></ul><ul><li>Characteristic hooked ends </li></ul><ul><li>Leptos-thin </li></ul><ul><li>L interrogans-pathogenic-divided into several sero groups </li></ul>
  28. 33. Field mice F,lymphade Sevenday fever Hebdomadis pig Fver d/s Swineherd’s Pomona Field mice Fever,prostration,aseptic meningitis Swamp/marsh fever Grippotyphosa Dog Influenza like,aseptic meningitis Canicola fever canicola rat Fever,jaundice,haemorrhage Weil’s d/s Icterohaemorrhagiae Animal reservoir CF Disease Serotype
  29. 34. Cattle F Dairy farmer fever Hardjo rat F Indonesian weil’s d/s Bataviae pig fever Febrile spirochetosis Pyrogenes ------ Fever rash over tibia Peritibial fevr/FBF Fortbragg
  30. 35. <ul><li>Morphology-leptospires are delicate flexible helical rods </li></ul><ul><li>Num. coils-so close together so can be diff. only under dark ground illumination </li></ul><ul><li>Ends are hooked resembles umbrella handles </li></ul><ul><li>Actively motile </li></ul><ul><li>Stain poorly with anilline dyes </li></ul><ul><li>Stained with Geimsa stain </li></ul><ul><li>Better results with silver impregnation methods </li></ul>
  31. 36. <ul><li>Culture-grown in media enriched with rabbit serum.use 5-fluro uracil to avoid contaminents </li></ul><ul><li>Simple method to avoid contaminents-inoculate intraperitoneally in guinea pigs and culture the heart blood collected 10 min later----lepts are able to invade the blood stream more rapidly than other bacteria </li></ul><ul><li>In natural host assymptomatic—accidentally reaches humans—d/s </li></ul><ul><li>Liquid and semisolid media-Korthof’s media,Stuart’s media,Fletcher’s media </li></ul><ul><li>Semisynthetic-EMJH </li></ul><ul><li>Growth ch. Few mm below the surface </li></ul>
  32. 37. <ul><li>Antigenic properties-exhibits considerable antigenic cross reaction. </li></ul><ul><li>A genus specific somatic antigen is present in all members of the genus </li></ul><ul><li>Pathogenicity-in natural reservoir assymptomatic </li></ul><ul><li>Infected urine—food –humans </li></ul><ul><li>Entes body thru cuts/intact mucosa of mouth, nose or conjunctiva </li></ul><ul><li>IP-10 days </li></ul>
  33. 38. <ul><li>Weil’s ds.-mild pyrexia ----fatal illness with hepatorenal damage </li></ul><ul><li>In severe cases the onset is acute-Rigor,vomiting ,headache,intense irritation of eye </li></ul><ul><li>Fever is irregular-subsides in about 10 days </li></ul><ul><li>Jaundice occurs in 10-20 % by seond or third day. </li></ul><ul><li>Purpuric he. May ccur in skin and mucosa </li></ul><ul><li>Albuminuria is a constant feature </li></ul>
  34. 39. <ul><li>Clinically presentation –Icteric and nonicteric </li></ul><ul><li>Many cases presents as aseptic meningitis </li></ul><ul><li>,sometimes abdominal symptoms predominate </li></ul><ul><li>Clinical dg. Is impossible in majority of cases </li></ul><ul><li>Leptosires are seen in blood in acute conditions, but not after 8-10 days </li></ul><ul><li>Persists in kidneys-demonstrated in urine in the later stages if the disease </li></ul>
  35. 40. <ul><li>Lab dig-demonstration micro.,culture ,inoculation,serology </li></ul><ul><li>Examination of blood-only in early stages-and b4 antibiotics given— </li></ul><ul><li>Dark field micro. </li></ul><ul><li>3-4 drops blood---EMJH----37C—2days—after tht room temp.—2weeks-------samples are examined on every third day under darkground for lept. </li></ul><ul><li>Primary isolation may be dalayed-may take weeeks to months- </li></ul>
  36. 41. <ul><li>Blood—I/perit. –guinea pigs-dies of fever and jaundice in 8-12 days (if icterohaemorrhagiae) ,others(canicola,pomona)-no effect---from 3 rd day-peritoneal fluid is examined daily </li></ul><ul><li>Blood from cardiac puncture is inoculated into cultural media </li></ul><ul><li>Examination of urine-appears in 2 nd week of ds. And intermittently thr after for 4-6wks </li></ul><ul><li>Examine immediately aftr voiding-as lysed in acidic urine/centrifuged deposits under dark field </li></ul>
  37. 42. <ul><li>Serological dg.-Antibodies appear in serum towards the end of the first week,and increase till the 4 th week,declining thr aftr. </li></ul><ul><li>Agglutinins are demostrable years after infection. </li></ul><ul><li>Two types f tests </li></ul><ul><li>Broadly reactive screening tests/genus specific tests </li></ul><ul><li>Serotype specific tests </li></ul>
  38. 43. <ul><li>Broadly Specific-antigens from L.biflexaPatoc 1 strain </li></ul><ul><li>Tests- </li></ul><ul><li>Sensitised erythrocyte lysis-(SEL) </li></ul><ul><li>CF </li></ul><ul><li>Agglutination </li></ul><ul><li>IF </li></ul><ul><li>Dip-stick assay </li></ul><ul><li>ELISA-detect IgM &IgG-to indicate the stage of the disease </li></ul>
  39. 44. <ul><li>Type specific tests-to identify the infecting serovar </li></ul><ul><li>Macrscopic Agglutination test-Formalinised Lpt.serovars * test serum---for agglutination –in serial dilns. </li></ul><ul><li>Microscopic Agglutination test(MAT)-live culture*test serum—in lowpower dark field—more specific </li></ul>
  40. 45. <ul><li>Diag thru animals-by culturing pieces of kidney </li></ul><ul><li>Ex. Of water-shaved and scarfified area of the skin of a guinea pig is immersed in water for an hour –infection take place thru abrasions </li></ul><ul><li>Px-Doxycycline 200mg.p.o. per week </li></ul><ul><li>T/t-if serious- </li></ul><ul><li>Penicillin-1-2million units-iv-6 th hourly -10 days </li></ul>