By R.Teja sri Introduction Coxiella burnetti is the causative agent of ‘Q-fever’ Obligate intracellular, gram negative bacterium Distributed Found globally in many species of animals Morphology:obligate intracellular pathogen . gram negative . Pleomorphic . size : rods:- 0.2 – 0.4 x 0.4 – 1.0 mc spheres :- 0.3 – 0.4 mc filterable . better stained with GIMINEZ and other rickettsiael stains . C. burnetii i en.wkipedia.org Culture Grows well in yolk sac of chick embryos and in various cell cultures . Ag structure shows phase variation . phase – I ,II . phase – I :- autoagglutinable more immunogenic activity due to periodate sensitive trichloracetic acid-soluble surface carbohydrate . o Phase – II :- more suitable for CFT . o both phase I ,II elicit good Ab response . Resistance Resistant to physical and chemical agents In pasteurization flash method is effective Can survive in dust and aerosols Inactivated by 2% formaldehyde 5% H2O2 1% Lysol . Contd…. Resistant to heat, drying and disinfectants Air samples test positive for 2+ weeks Soil samples test positive for 150+ days Spore formation PATHOGENESIS History Q stands for Query or Queensland Origin of disease unknown First reported cases were in Queensland, Australia Differentiating features : 1. Having smaller size 2. Resistance to heat and drying 3. Major route of transmission isinhalation/ingestion Primary Reservoir Goats Cattle Sheep * All eukaryotes can be infected Bacteria is excreted in: Feces Urine Milk of infected animals Release Into Environment: During birthing the organisms are shed in high numbers in amniotic fluids and the placenta 109 bacteria per gram of placenta Do not touch! Transmission Most common route is inhalation of aerosols Contaminated dust, manure, birthing products Tick bites (rare) Human to human also very rare gsbs.utmb.edu Contd….. Who’s at risk? Farmers, veterinarians, researchers, abattoir (slaughterhouse) workers etc. People who breed animals Immunocompromised Acute or Chronic Q fever gsbs.utmb.edu *Bacteria spread through blood Symptoms Acute Q fever Self-limiting, flu-like disease Fever, nausea, headaches, vomiting, chest/abdominal pain Pneumonia & granulomatous hepatitis Chronic Q fever (> 6 months) Endocarditis & meningoencephalitis Pre-existing disease Host interaction Entry via inhalation Alveolar macrophages encounter bacteria C. burnetii phagocytosed Macrophage C. burnetii R Heinzen, NIAID Host interaction Replication Low within phagolysosme pH needed for metabolism No cellular damage unless lyses occurs Can invade deeper tissue and cause complications Phagocytosis Binding/entry into macrophages via: Integrin Associated Protein (IAP) Leukocyte Response Integrin (LRI) macrophage bacteria Binding & Entry Phagocytosis Phagocytic vesicle Lysis of phagolysosome and macrophage Phago-lysosome fusion: bacteria survive and multiplies LAB DIAGNOSIS Hard to diagnose because: Asymptomatic in most cases Looks like other disease (Flu or cold) Serology continues to be best method PCR, ELISA and other methods WEIL – FELIX test is negative . Contd….. Bio safety level 3 (BSL-3) facility Very infectious (one organism causes infection) Listed by the CDC as a potential bioterrorism agent. Isolated in cell cultures or embryonated eggs Treatment Once infected, humans can have life-long immunity Acute Q fever treated with: doxycycline, chloramphenicol, erythromycin or fluoroquinolones Chronic Q fever treated with: Vaccines : prepared from formalin killed whole cells attenuated strains trichloroacetic acid extracts Prophylaxis:Pasteurization and sterilization of milk and other dairy products Disinfect utensils, machines used in farm areas for birthing Regular testing of animals and those who work closely with them Protective Personal Equipment BARTONELLA INTRODUCTION Family Bartonellaceae contain two genera Bartonella Grahamella Grahamella does not infect humans Bartonella contain 3 species: B.bacilliformis B.quintana B.henselae BARTONELLA BACILLEFORMIS Carrions disease Causes OROYA fever MORPHOLOGY: Gram negative Pleomorphic strict aerobe motile, small bacillu0.3-0.5x0.2-0.5mc found inside erytrocyte infected persons Opt. temp 25-28 c CULTURE; Grow in semisolid nutrient agar with 10% rabbit serum 0.5%Hb Growth is slow takes about 10 days PATHOGENISIS: Causes OROYA fever Transmitted by SAND flies INCUBATION PERIOD; 3 weeks to 3 months CLINICAL FEATURES: Fever Headache Chills Severe anemia Several weeks after recovery pt. develop nodular lesions on the body Secondarily infect produce ulcers – VERUGA PERUANA Lab diagnosis:Demonstrated in blood smear by GIEMSA stain Seen in cytoplasm and adhere to cell surface Grown on NA agar contain rabbit serum, Hb Guinea pig inoculation leads to VERUGA PERUANA TRETMENT: Susceptible to penicillin streptomycin Tetracycline Chloramphenicol PREVENTION Insecticides such as DDT should be used to eliminate sand flies BARTONELLA QUINTANA MORPHOLOGY:small gram negative bacillus 0.3-0.5 mc to1.0-1.7 mc Does not posses flagella show twitching movments by fimbriae CULTURE: Grows on rabbit /sheep blood agar opt. temp -35 c in 5% CO2 colonies appear after 14 days primary inoculation PATHOGENESIS:Formerly called Rochalimaea quintana Causes TRENCH fever also called FIVE DAY fever Transmission; by body louse vertical transmission does not occur in lice Lice after acquiring infection remain infectious through out life CLINICAL FEATURES: Mild symptoms leads to chronic rickttesiaemia Relapse have been observed even after 20 years primary disease Lab diagnosis: Detected in the gut of infected lice Isolate from pt. blood by cultur sheep blood agar Weil-felix test negative PCR- detect organism in tissues BARTONELLA HENSELAE MORPHOLOGY:Gram negative Slightly curved Show twitching movments CULTURE:Grows on chocolate agar columbia agar with 5%sheep blood tryptic-soy agar opt.temp-35-37 c in 5% CO2 COLONY MORPHOLOGY:- white, dry, cauliflower like and embedded in the agar PATHOGENESIS: Causes CAT-SCRATCH disease Occur by contact with scratch / bite of an infected cat Resolution in weeks to months 1 - 3 weeks Cat contact (scratch, bite, ? cat flea bite) Dissemination in immunocompromised hosts CLINICAL FEATURES: Regional lymphadenopathy Fever Endocarditis In AIDS pt. leads to; bacillary angiomatosis Lab diagnosis: lymph node biopsy – stained with WARTIN-STARRY SILVER IMPREGNATION –clusters of bacillus Grow on chocolate agar/ columbia agar TREATMENT: Self limiting No specific treatment required