1. Pathogenic spirochetes. Rickettsia. Mycoplasmas. Clamidia. Chair of Medical Biology, Microbiology, Virology, and Immunology Lecturer As. Prof. O.V. Pokryshko

2. Spirochetes Treponema Borrelia Leptospira

3. Main pathogenic bacteria of Spirochetaceae GenusSpeciesSubspeciesDisease Treponema T.pallidum T.сarateum T.vincentii pallidum endemicum Pertenue Syphilis Bejel Yaws Pinta Vincent’s angina Borellia B.recurrentis B.caucasica, B.duttoni, B.persica та ін Epidemic relapcing fever Endemic relapsing fever B.burgdorferiLyme disease Leptospira Icterohaemo- rrhagiae Leptospirosis

4. Spirochetes structure

5. Spirochetes morphology Treponema Leptospira Borrelia

6. Spirochetes ultrastructure Borrelia Leptospira

7. Classification of Human Terponema  Non-pathogenic T.denticola T.macrodenticum T.orale  Conditionally pathogenic T.vincentii  Pathogenic T.pallidum pallidum (syphilis) T.pallidum endemicum (bejel) T.pallidum pertenue (yaws) T.carateum (pinta)

8. Vincent’s angina

9. Noma (gangrenous stomatitis)

10. Treponema in tested material

11. Cell attacked by treponema

12. Hard chancre

13. Clinical findings of Syphilis Secondary syphilis Hetchinzone teeth

14. Microbiologic diagnosis of Syphilis  Microscopy (native an fixed material) Romanovsky-Giemsa stain Phase contrast Dark field  Serologic diagnosis Wassermann’s test (CFT) Sedimentation reartions (Каhn’s and Sachs- Witebsky’s tests) Microreaction of Treponema pallidum immobilization IFT, ELYSA, IHAT Chain polymerase reaction

15. Tropic trepanematoses: pinta (T.carateum), bejel (T.endemicum), yaws (T.pertenue) Yaws (tropic granuloma) Bejel (endemic syphilis)

16. Yaws

17. Pinta

18. Bejel

19. Borrelia morphology

20. Borrelia in blood smear

21. Lyme disease Causative agents: Borrelia burgdorferi Borrelia garinii Borrelia afzelii

22. B. burgdorferi

23. Leptospira morphology

24. Clinical findings of leptospirosis

25. Rickettsia

26. General Features The rickettsia are bacteria which are obligate intracellular parasites. They are considered a separate group of bacteria because they have the common feature of being spread by arthropod vectors (lice, fleas, mites and ticks). The cells are extremely small (0.25 u in diameter) rod-shaped, coccoid and often pleomorphic microorganisms which have typical bacterial cell walls, no flagella, are gram-negative and multiply via binary fission only inside host cells. They occur singly, in pairs, or in strands.

27. General Features They occur singly, in pairs, or in strands. Most species are found only in the cytoplasm of host cells, but those which cause spotted fevers multiply in nuclei as well as in cytoplasm. In the laboratory, they may be cultivated in living tissues such as embryonated chicken eggs or vertebrate cell cultures.

28. The family Rickettsiaceae is taxonomically divided into three genera:  1. Rickettsia (11 species)--obligate intracellular parasites which do not multiply within vacuoles and do not parasitize white blood cells.  2. Ehrlichia (2 species) – obligate intracellular parasites which do not multiply within vacuoles but do parasitize white blood cells.  3. Coxiella (1 species) – obligate intracellular parasite which grows preferentially in vacuoles of host cells.  4. Bartonella (3 species) – intracellular parasite which attacks the red blood cell.

29. Structure The structure of the typical rickettsia is very similar to that of Gram-negative bacteria. The typical envelope consists of three major layers: an innermost cytoplasmic membrane, a thin electron dense rigid cell wall and an outer layer. The outer layer resembles typical membranes in its chemical composition and its trilaminar appearance. The cell wall is chemically similar to that of Gram-negative bacteria in that it contains diaminopimelic acid and lacks teichoic acid. Intracytoplasmic invaginations of the plasma membrane (mesosomes) and ribosomes are also seen. There are no discrete nuclear structures

30. Pathogenicity In their arthropod vectors, the rickettsia multiply in the epithelium of the intestinal tract; they are excreted in the feces, but occasionally gain access to the arthropods salivary glands. They are transmitted to man, via the arthropod saliva, through a bite. In their mammalian host, they are found principally in the endothelium of the small blood vessels, particularly in those of the brain, skin and heart.

31. Pathogenicity Hyperplasia of endothelial cells and localized thrombus formation lead to obstruction of blood flow, with escape of RBC's into the surrounding tissue. Inflammatory cells also accumulate about affected segments of blood vessels. This angiitis appears to account for some of the more prominent clinical manifestations, such as petechial rash, stupor and terminal shock. Death is ascribed to damage of endothelial cells, resulting in leakage of plasma, decrease in blood volume, and shock.

32. Pathogenicity It is assumed that the observed clinical manifestations of a rickettsial infection are due to production of an endotoxin, although this endotoxin is quite different in physiological effects from that produced by members of the Enterobacteriaceae.

33. Laboratory Diagnosis Presumptive laboratory diagnosis is based on the finding of rickettsial-like organisms in tissue or blood. Although the organisms are gram-negative, they only weakly take the counter stain, safranin. Therefore, special staining procedures are used. Infected tissue may be stained with:  1. Macchiavello stain--organisms are bright red against the blue background of the tissue.  2. Castaneda stain--blue organisms against a red background.  3. Giemsa stain--bluish purple organisms.

34. Laboratory Diagnosis Confirmative diagnosis is based on a serological reaction (Weil- Felix reaction) in which the titer of the agglutinins in the patient's serum against the Proteus strains OX-19, OX-2 and OX-K are determined. These Proteus strains have no etiological role in rickettsial infections, but appear to share antigens in common with certain rickettsia. These antigens are alkali stable polysaccharide haptens which are distinct from the group-specific and type-specific antigens. In interpreting the results, it must be kept in mind that Proteus infections are fairly common (especially in the urinary tract) and that they, too, may evoke antibodies to the Proteus-OX strains. This test is usually positive seven days after the initial infection.

35. Laboratory Diagnosis A more specific complement fixation test is available but does not show positive results until 14 days into the infection. The indirectfluorescent antibody test is also useful for the detection of IgM and IgG antibodies against rickettsia. In fact, this is the diagnostic test of choice for ehrlichiosis.

36. Diseases 1. Louse-borne: European epidemic typhus (Rickettsia prowazekii), Brill's disease (Rickettsia prowazekii), Trench fever (Bartonella quintana) 2. Flea-borne Endemic murine typhus (Rickettsia typhi), Cat scratch fever /Bacilliary angiomatosis/ (Bartonella henselae) 3. Mite-borne Scrub typhus (Orientia /Rickettsia tsutsugamushi), Rickettsialpox (Rickettsia akari)

37. Diseases 4. Tick-borne Rocky Mountain Spotted Fever (Rickettsia rickettsii), North Asian tick typhus (Rickettsia siberica), Fievre boutonneuse (Rickettsia conorii), Queensland tick typhus (Rickettsia australis), Q-fever (Coxiella burnetii), Spotted fever (Rickettsia rhipicephali), Ehrlichiosis (Ehrlichia canis, Ehrlichia chaffeensis) 5. Fly-borne Oroyo fever / Verruga peruana (Bartonella bacilliformis)

38. Chemotherapy The drugs of choice for the treatment of rickettsial diseases are chloramphenicol and tetracycline. Each of these is highly toxic, especially in children, and must be used with care. The sulfonamides stimulate rickettsial growth and thus are contraindicated in the treatment of these diseases.