Diagnostic microbiology lecture: 17 CHLAMYDIA Abed ElKader Elottol MSc . Microbiology 2010

1. Diagnostic microbiologylecture: 17CHLAMYDIA Abed ElKaderElottolMSc. Microbiology 2010 Abed ElKaderElOttol

2. General Characteristics • Obligate intracellular bacteria. • Have ribosomes like bacteria. • Are metabolically deficient. Morphology • Small rounded organism. • Multiply by binary fission. • Cell wall consists of inner & outer membranes but differ from that of Gram negative bacteria by absence of peptidoglycan. Abed ElKader ElOttol

3. Important Species • C. psittaci • C. pneumoniae • C. trachomatis • Cultural Characters Grow in: • Yolk sac of chicken embryo • Tissue culture (McCoy cells) • PATHOGENESIS • • Infect epithelial cells of mucous membranes & lungs • Virulence is due to: • • Resistance to phagocytic killing by lysosomal enzymes • • Heat-labile toxin • • Competition with host cell for nutrients • • Host’s immune response may account for inflammation & tissue destruction. Abed ElKader ElOttol

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5. CHLAMYDIA PSITTACI • A zoonotic respiratory disease. • Natural habitat : birds • Transmitted through inhalation of : • Respiratory secretions & dust from faeces of infected birds • Common in poultry workers • Disease : Pneumonia (Psittacosis) • Diagnosis • Isolation of organism from sputum by tissue culture • Complement fixation test to detect specific Abs • Treatment • Tetracycline in adults • Erythromycin in babies Abed ElKader ElOttol

6. CHLAMYDIA PNEUMONIAE Also known as TWAR (TW – Taiwan & AR – acute respiratory) Cause atypical pneumonia like Mycoplasmapneumoniae Treatment Tetracycline in adults Erythromycin in babies Abed ElKader ElOttol

7. CHLAMYDIA TRACHOAMATIS 15 serotypes (A-L) Transmission: Through close personal contact like: • Sexual • Passage through birth canal • Finger to eye or fomite to eye (Trachoma) DISEASES 1. Trachoma • Caused by serotypes A, B, Ba & C • One of the leading causes of blindness in developing countries with dry & hot weather • Chronic conjunctivitis : leads to scarring of eye lids and cornea Abed ElKader ElOttol

8. 2. Genital Tract Infections (Serotypes D-K) • Non-gonococcalurethritis in men • A common cause of non-gonococcalurethritis • Mucopurrulent urethral discharge • May progress to epidydmitis & orchitis (testes inflamation) • Cervicitis & Vaginitis • Mucopurrulent vaginal discharge • Pelvic Inflammatory Disease (PID) • May lead to secondary infertility Abed ElKader ElOttol

9. 3. Neonatal Infections (Caused by serotypes D-K) • Acquired from mother’s birth canal • Inclusion Conjunctivitis • Profuse mucopurrulent discharge 7-12 days after birth • Pneumonia. 4. LymphogranulomaVenereum (LGV) • Caused by serotypes L1, L2 & L3 • A STD with lesions on genitalia & LNs (buboes) 5. Reiter’s Disease An autoimmune disease caused by Abs formed against C. trachomatis which cross react with antigens on cells of urethra & joints(arthritis, redness of the eyes, and urinary tract signs). Abed ElKader ElOttol

10. LAB DIAGNOSIS • • Specimens from urethra, conjunctiva, sputum & cervix. • • Microscopy. • • Chlamydial “cytoplasmic inclusions” are detected by: • • Giemsa staining • • Fluorescent Ab staining • • PCR • • Cell Culture • • Sero-diagnosis • TREATMENT • Tetracycline in adults • Erythromycin or Azithromycin in babies Abed ElKader ElOttol

11. Calymmatobacteriumgranulomatous GRANULOMA INGUINALE Abed ElKader ElOttol

12. The disease is commonly known as donovanosis • Small, painless nodules appear after about 10–40 days of the contact with the bacteria. Later the nodules burst, creating open, fleshy سميك, oozingرشح lesions. • General characteristics • Capsulated short Gram-negative rod • A STD with higher incidence in homosexuals • Clinical Features • Initially papules appear on external genitalia which ulcerate and extend widely – ulcer formation • Base of ulcer is “BEEFY”; spreads by contact so is known as “KISSING ulcers” • Lymph Nodes may enlarge • Treatment : Tetracycline Abed ElKader ElOttol

13. MYCOPLASMA Abed ElKader ElOttol

14. GENERAL CHARACTERISTICS • The smallest free-living organism (0.3 μ diameter). • Have no cell wall. • Insensitive to penicillins & cephalosporins. • Poorly stained by Gram-staining. • Cytoplasmic membrane contains cholesterol. • Slow growth on specialized artificial culture media (a week). • Typical “fried-egg” appearance of colonies by a plate microscope Abed ElKader ElOttol

15. Formulae Difco™ PPLO Agar Approximate Formula* Per Liter Beef Heart, Infusion from 50 g ................................... 6.0 g Peptone ................................................................... 10.0 g Sodium Chloride ........................................................ 5.0 g Agar ......................................................................... 14.0 g Abed ElKader ElOttol

16. MYCOPLASMA PNEUMONIAE MAIN DISEASE • Primary atypical pneumonia • Common in late summer and early autumn PATHOGENESIS & EPIDEMIOLOGY • Droplet infection. • Organism adhere to respiratory epithelium. • Inhibit ciliarymotion. • Damage epithelium. • Common in chidren & young adults • Increased incidence in winter Abed ElKader ElOttol

17. CLINICAL FEATURES • Sore throat, fever & headache. • Cough with small amount of whitish non-purulent sputum. • Some extrapulmonarysymptoms. • Opacities on chest X-Rays. • IMMUNITY • Incomplete: second episode can occur. Abed ElKader ElOttol

18. UREAPLASMA URELYTICUM Abed ElKader ElOttol

19. Differentiated from mycoplasma due to urease enzyme production Like mycoplasma produce “fried egg” colonies on specialized medium. • Diseases • • Non-gonococcal, non-chlamydial urethritis in men. • • Post-partum fever in women. • • Transmitted by sexual contact. • MYCOPLASMA & UREPLASMA • LAB DIAGNOSIS • Culture : “Fried egg” colonies on specialized medium • Cold Agglutinin detection A titer of 1:128 or higher – indicates recent infection • TREATMENT • Tetracycline OR Spectinomycin Abed ElKader ElOttol

20. RICKETTSIA & COXIELLA Abed ElKader ElOttol

21. Rickettsiae • A genus of small, rod-shaped, round to pleomorphic microorganisms . • gram-negative • cultivable only in living tissues. • Transmitted by liceقمل and ticksقراد, they cause disease in humans and domestic animals but are also found in the cytoplasm of tissue cells of lice, fleas, ticks and mitesسوس , which may act as reservoirs and vectors

22. Transmission • Biological • Examples • Ticks • Sucking lice • Live inside arthropods for months • Mechanical • Examples • Instruments • Blood transfusions • Live short time outside of host

23. Pathogenesis • Insect vectors for human transmission: arthropods such as fleas, ticks, mites عث, or lice • Rodents, humans, or arthropods: all can serve as reservoirs • Multiply in salivary glands of ticks & gut of lice/fleas • Site of Bite = Eschar (encrustedمتقشرulcer) → necrotic tissue:

24. Site of bite = point at which Rickettsia enters the body becomes blackened • Following bite, organisms taken into body by “phagocytosis process • Organisms multiply in both nucleus & cytoplasm of host cell.

25. Typhus Group • Louse borne (epidemic) Typhus. • Etiological agent = Rickettsiaprowazekii • Transmission: person-to-person by infected human body louse that excretes organisms in feces • Introduction of pathogen form lice facilitated by scratching the louse bites • high fever, chills, rash possible • Duration of Disease: ~2 weeks; more severe in ederly • Complications: CNS dysfunction and myocarditis

26. Rock Mountain Spotted Fever • Etiological agent = Rickettsiaricketsii • Reservoir = rodents (mice, rats) • Vector = Tick Bite • Rickettsiain saliva of tick, transfer to humans after bite Human infection • High fever & malaise, then rash – initially, but becomes petechial or hemorrhagic • Rash starts on extremities, then rapid spread to entire body. • Untreated cases – vascular disturbances with myocardial or renal failure possible

27. Coxiella • Etiological agent = Coxiellaburnetti • Q fever • Reservoir = cattle, sheep, rodents, ticks • Vector = contaminated aerosol • C. burnettienters the body via mucous membranes, abrasions & GIT via consumption of milk for infected animals • Zoonosisbecouseanimal direclty transmits the organism to humans • high fever, cough, pneumonia, hepatitis – all self-limiting usually; rarely – endocarditis (scarring of hrt. valves) and scarring of liver • complicated by hepatitis, myocarditis, or encephalitis

28. Growth characteristics • Grow only in eukaryotic cell like • Tissue cultures OR embryonated eggs • Abed ElKader ElOttol

29. RICKETTSIA & COXIELLA : LAB DIAGNOSIS • Serology • PCR • Tissue culture • Weil-Felix Reaction • Antigens of several species of Rickettsiae cross-react with cell wall O antigen of Proteus OX-2, OX-19, OX-K. • These Proteus antigens can be used in lab to detect presence of specific antibodies against certain Rickettsia in patients serum. • Reaction negative in Q fever TREATMENT • Tetracycline • Chloramphenicol Abed ElKader ElOttol

30. End of Lecture Abed ElKaderElOttol