Microbiology

Bacteria Morphology Streptococcus Pneumoniae Pneumococcus -Gram +ve diplococcic, lanceolated with distal ends narrowed -Non sporing, possess a capsule that permit typing with specific antisera Moraxella Catarrhalis -Opportunistic pathogen -Gram ve diplococci Corynebacterium Diphtheria -Gram +ve bacilli -Form sharp angle to each other resembling Chinese letter writings -Clubbing at one or both ends -Contain volutin metachromatic granules which are intracellular depots of polyphosphate and give the rod a beaded appearance. Better seen when stained by Neissers method -Better demonstrated when the film is taken from culture grown on Loefflers serum medium Specimen: Throat swab Smears: 1)Grams stain -Gram +ve bacilli, -ve result is common -Vincents angina: numerous fusiform bacilli and Borrelia vincenti are seen. The spirochaetes only appear if dilute carbol fuchsin is used instead of safranin as counterstain -Candidal oral thruth: Gram +ve budding oval bodies -Acute tonsillitis: Gram +ve cocci in chain 2)Neissers stain -to detect metachromatic granules Culture: -Loefflers serum are examined by Gram stain -Blood agar plate are examined, if -haemolyitc, maybe streptococcus pyogenes or rarely mixed with diphtheria -Mcleods tellurite plate is inhibits other inhabitants, the black colonies are isolated to be cultured on Loefflers slope to be identified by Gram stain and tested for toxin production Toxigenicity of virulence tests: 1)Eleks test

Laboratory Diagnosis

Specimen: 1) Blood (bacteremia) 2) CSF (meningitis) 3) Sputum (lobar / bronchopneumonia) Culture: -Aerobe and facultative anaerobe -37oC -No grow on MacConkeys agar or nutrient agar but on media containing 5-10% serum or blood -Colonies on blood agar are small, smooth, transparent, low convex and -haemolytic -With prolonged incubation, the colonies become depressed or flattened centrally due to autolysis -Must be differentiated from streptococcus viridians by optochin test, bile solubility, inulin fermentation, virulent in mice

Created by Qosru Iskandariah

Antigenic Structure

-83 serotypes are distinguished by different in polysaccharide antigen present in capsule -Determined by reaction with specific antisera (capsular swelling reaction) -Commonest bacterial pathogen in lobar or bronchopneumonia -Complications: 1) Pleurisy 2) Emphysema 3) Meningitis 4) Arthritis 5) Sinusitis 6) Brain abscess -Produce disease by multiplying inside tissue -Capsule inhibits phagocytosis 1) Penicillin 2) If resistant, vancomycin -Pneumonia -Sinusitis -Otitis media

Pathogenesis

-A strip of filter paper saturated with antitoxin is placed on a serum agar plate -The strain tested is streaked at right angle to the filter paper and incubated at 37oC for 2 days -The antitoxin diffusing from the strip with form precipitate lines with toxin from toxigenic strain -Absence of precipitate line indicates non toxigenic 2)Enzyme-linked immunosorbent assays -To detect diphtheria toxin 3)PCR -To detect diphtheria toxin gene -Diphtheria toxin are powerful exotoxin that inhibits protein synthesis -Lysogenized strain of diphtheria are toxigenic and virulent, while non lysogenized strain are non toxigenic and avirulent -In the soft part of faces, or may infect wounds, conjunctiva, vulva -At the part affected, pseudomembrane is produced with a numerous diphtheria bacilli -Localized and no invasion to blood stream -Symptoms include paralysis and myocardium degeneration due to exotoxin that has special affinity to heart muscle, adrenal cells and nerve cells -Incubation period is 2-5 days

Treatment

Prophylaxis

-Polyvalent vaccine

1)Diphtheria antitoxic serum -IM or IV -Skin test for sensitivity to horse serum before treatment -If reaction occurs (erythema in 20min), desensitization is done 2) Antibiotics (penicillin and erhthromycin) -To reduce number and stop secretion of exotoxin 1)Toxoid (active immunization) 2) DPT for children under 6 years, and above without pertussis vaccine

Created by Qosru Iskandariah

Haemophilus
Bacteria Morphology Haemophilus Influenzae -Small Gram ve coccobacilli and rods -Pleomorphism -Capsules are present in isolated recently organisms in young culture -Capsules are divided into 6 serological types according to nature of polysaccharide -Type b is major virulence antigen -Antiphagocytotic -Lipooligosaccharides are endotoxin -Acute epigottitis -Otitis media -Pneumonia The non typable strains responsible to: -Acute sinusitis -Chronic bronchitis, bronchiectasis, purulent sputum Bordetella Pertussis -Gram ve coccobacilli -Capsulated, non motile Legionella Pneumophila -Gram ve short rods -Stain poorly and gram stain smears from clinical specimen are ve -Serogroup 1 is the most common group pathogen

Antigenic Structure

-Haemolytic -Produce pertussis toxin, tracheal toxin, dermnecrotic toxin -Has pili for adherence -Whooping cough -Transmission by droplet infection -Incubation period about 2 weeks: 1) Catarrhal stage: Mild coughing, sneezing, numerous bacteria are expelled in drops 2) Paroxysmal stage: Explosive whooping cough, vomiting, cyanosis, convulsion

Pathogenesis

Laboratory Diagnosis

Specimen: -Nasopharyngeal swab -Blood -CSF -Pus Smear: -If the organisms are present in large number, can be detected by Gram stain Culture: -Facultative anaerobe and aerobes -37oC -Not grow in ordinary media, require certain

Specimen: -Pernasal swab (best efficient) -Nasopharyngeal swab -Cough plate (least efficient) Culture: -Aerobe -Primary isolation is facilitated by enriched media (Bordet Gengou plates/charcoal yeast extract agar + cephalexin) -Inoculate on Bordet Gengou plates, one with or without penicillin -Incubate at 37oC for 3-7 days in a moist condition -Colonies are identified by slide agglutination with

-Asymptomatic infection -Clinically significant disease during old age, debilitated, immunocompromised patients -Infection through air -Most serious infection is lobar pneumonia (Legionnaires disease) -Legionella enters and grows within human macrophage and monocytes and not efficiently killed by polymorphs, they multiply within until autolysis occurs and the released bacteria infect other macrophages -Not communicable Specimens: -Bronchial washings -Pleural fluid -Lung biopsy -Blood Smear: -Not demonstrable in gram stain -Direct fluorescence antibody can be done Culture: -Fastidious -Grow on complex media (Buffered Charcoal Yeast Extract agar, BCYE) at pH 6.9 at 35oC

Created by Qosru Iskandariah

growth factors: X factor (Haemin) and V factor (NAD) -Blood contain X factor and RBC contain V factor -Grow in chocolate agar but better growth in chocolate agar + isovilatex (mixture of NAD and other growth factors)

Bordetella pertussis antiserum or immunofluorescence stain PCR (most sensitive method using primers)

and humidity 90% -Grow slowly after 3 days -Pleomorphism -Catalase and oxidase +ve Serology: -Antibody level rises slowly, so useful in retrospective study of outbreak Immunity: Both humoral and cell-mediated immune response occur but cell-mediated are more important due to intracellular infection -Erythromycin

Treatment

Prophylaxis

-Ampicillin, but 25% are resistant due to B lactamase production -New cephalosporins -H influenza conjugate vaccine (H influenza type b polysaccharide vaccine + protein carrier as mutant diphtheria toxin protein or toxoid) -Rifampin (children less than 4 years)

-In catarrhal stage: erythromycin as prophylaxis -In paroxysms stage: antibiotic cannt cure -DPT (children under 6 years) -Acellular vaccine (contain 1-5 antigens) can be given to children above 6 years

-Cleaning, chlorination and heating of water

Created by Qosru Iskandariah