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Chlamydiacae The taxonomy of Chlamydiacae has been revised on the basis of genomic studies; and accordingly they have been divided into 2 genera: Chlamydia Chlamydia trachomatis 2 biovars Trachomatis & LGV Chlamydophila Chlamydia psittaci Chlamydia pneumoniae They were once considered as viruses because : They are small enough to pass through o.45µ filters They are obligate intracellular parasites. They are now considered as bacteria because: 1.They have inner & outer membranes similar to gram negative bacteria. 2. They contain both DNA &RNA 3.They can synthesize their own proteins, nucleic acids & lipids 4.They are susceptible to many antibiotics. Antigenic structure 1. They have a genus-specific lipopolysaccharide detected by complement fixation test. 2. They have species &strains-specific outer membrane proteins Staining 1. Giemsa………….stains the elementary bodies , the reticulate bodies &inclusions (not for definitive diagnosis ) 2. Gram……………gram negative or gram variable (difficult ) 3. Immunofluorescense 4. Iodine……………for intracellular inclusions which contain glycogen Developmental cycle of Chlamydia -EB (elementary body ) attaches to the surface of susceptible cell & enters the cell by phagocytosis -The elementary body organizes into RB ( Reticulate body ). -The reticulate body divides by binary fission. -After 24-48 hrs ,EBs are released and initiate a new cycle of infection -The mass of EBs → Inclusion body→detected by histologic stains NB 1-After internalization,bacteria remain within the cytoplamic phagosome & replicate. 2-Fusion of cellular lysosomes & EBs containing phagosome , and subsequent intracellular killing is inhibited (bacteria not affected by lysosymes Growth Eukaryotic cell lines : Hela cells-229 , Mc Coy cells , BHK -21 , Buffalo green monkey kidney cells. Sensitivity is increased by pretreatment with cycloheximide (to decrease host metabolism ), use of shell vial technique ( growth of host cell monolayer on glass cover slips rather than in small microtiter plates), use of Iodine stain or Fluorescein-conjugated antibodies to detect intracellular inclusions. - Embryonated egg yolk sac. - Mice (rarely used ) Reaction to physical & chemical agents Heat …………….at 60°C,for 10 min leads to their inactivation Ether……………..for 30 min………..leads to rapid inactivation Phenol 0.5%, for 24h…………………leads to inactivation Freeze drying…………………………decreases their infectivity Dryness……………………………….does not affect infectivity Treatment - Both sex-parteners should be simultaneously treated - Tetracyclins are commonly used in non-gonococcal urethritis and in non-pregnant females. - Azithromycin is also effective. - Erythromycin may be an alternative in pregnant females - Topical Tetracyclin or Erythromycin………for inclusion conjunctivitis. - In LGV……….Sulfonamides & Tetracyclins for the early stages;but late stages require surgery. Chlamydia Trachomatis It has a very limited range of infection (infects humans only) It has 2 Biovars: Trachoma (15 serovars A,B,Ba,C,D-K ) LGV (4 serovars L1,L2,L2a,L3) Clinical syndromes 1. Infections in Adults Non-gonococcal urethritis (NGU) in males - 50% of cases of NGU are sexually acquired. - 25% are asymptomatic but are able to transmit the organism. - When symptoms occur (urethral discharge,difficult micturition),they are mild (unlike gonococcal urethritis).Serious complications are rare. Mucopurulent cervicitis in females - It is the female counterpart of male NGU - It is acquired through sexual intercourse.Many remain asymptomatic. - The Gram stain of the endocervical swab shows yellow-green mucous and more than 10 PNLs/ HPF.(Neisseria must be excluded) - Complications include PID Pelvic inflammatory disease (PID) - It is an ascending infection. - Although symptoms may be mild yet laparoscopy may show severe inflammation. - Complications include salpingitis, endometritis,peritonitis, Prihepatitis(Fitz-Hugh Curtis syndrome).These may lead to infertility,chronic pelviabdominal pain & ectopic pregnancy. Lymphogranuloma venereum - It is a sexually transmitted disease. - The IP is about 4w. - The primary lesion occurs at the site of infection:vesicle,papule or ulcer,small,painless heals rapidly so it might be overlooked. - The second stage which occurs after2-5w shows marked inflammation& swelling of the lymph nodes (usually inguinal) - There is constitutional symptoms (usually severe).Fistulae may form (especially after needle aspiration) Acute urethral syndrome Occurs in young women in the form of recurrent dysuria,pyuria& sterile culture Ocular infections 1- Trachoma :( A,B,Ba,C ) keratoconjunctivitis,invasion of blood vessels into the cornea,bacterial infection&scarring. 2- Inclusion conjunctivitis :( A,B,Ba,D-K) in sexually active adults. It may occur as an autoinfection. Proctocolitis &epididymitis Reiter' s syndrome: conjunctivitis,reactive arthritis and urethritis. 2. Infections in infants Newborns………..from infected birth canal Infants pneumonia (1-6 mo ) : usually associated with conjunctivitis. Infants conjunctivitis :It is the commonest cause of neonatal conjunctivitis& is associated with mucopurulent discharge(2-3w after birth).Most cases resolve without sequelae. However,some may develop chronic ocular infection Diagnosis 1. Culture 2. Non-cultural methods: - Cytology : cell scrapings for inclusions,but is insensitive compared to culture &immunofluorescence. - Antigen detection: by direct immunofluorescence, than culture) ELISA(less sensitive - Nucleic acid probes: test the presence of a specific species-specific sequence of 16S rRNA.It is rapid & relatively inexpensive. - PCR,LCR,TMA (transcription mediated amplification), SDA (standard displacement ).They have a sensitivity of 90-98% In the very near future,they will be the test of choice. - Serology: has a limited value in Chlamydia trachomatis causing genital infections in adults,because antibody titers persist for a long period so,do not differentiate between concurrent and past infections; although a significant rise in antibody titer is useful. Chlamydia Psittaci Causes Psittachosis, Ornithosis, Parrot fever Humans are infected by contact with birds, inhalation of dried bird excrement, urine or resp. secretions. IP 4d C/ P: From mild inapparent or flu like inf. to severe pneumonia with sepsis and high mortality rate (20%) now decreased to 2%. Path: RT lungs. Blood Liver, Spleen, Kidneys and Diagnosis 1) Serology: 4 fold rise by CFT confirm by MIF Sometimes specific IgM Antibody can be demonstrated. 2) Cell culture: rarely performed Treatment Te, Macrolides Chlamydia Pneumoniae Was 1st isolated from conj. of a child in Taiwan (TW-183) and was found to be related to a pharyngeal isolate (AR-39) TWAR C. pneumoniae Chlamyolophila ( only a single serotype) Transmitted by resp. secretions (person to person) Human pathogen Common in adults Clinical Picture Usually mild or asymptomatic May cause bronchitis, pneumonia, sinusitis Cannot be diff. from other atypical pneumonias (Mycopl, Legionella,….) Associated with atherosclerosis Diagnosis difficult Do not grow Amplification techniques √ Serology: Complement Fixation: not specific (positive for both Chlamydia and Chlamydophia) IF √√ : the most sensitive and specific. It uses EBs as antigens Treatment E, Te, Lev 10-14d Characteristics of the Chlamydiae C. Trach. C. Pneum. C. Psittaci Inclusion Morphology Round, Vaeuolar Round, Dense Large, Variable Shape, Dense Glycogen in Inclu. Yes No No E B Morph. Round Pear Shaped Round Suspect. To Sulph. Yes No No Serovars 19 Natural Host Humans Mode of Transmission Person to person Mother to infant Trachoma STDs Infant pneumonia LGV Major Disease 1 Humans Air borne Person to person Pneumonia Bronchitis Pharyngitis Sinusitis ≥4 Birds Air borne (bird excreta to human ) Psittacosis Pneumonia Fever of U.O