1. Scientific classification Domain:Bacteria Kingdom:Eubacteria Phylum:ProteobacteriaClass:GammaproteobacteriaOrder:PasteurellalesFamily:PasteurellaceaeGenus:HaemophilusSpecies:H. influenzae

2. Medical important species Haemophilus influenzae Haemophilus aegyptius Haemophilus ducreyi

3. Normal habitatH.influenzae (mostly non-capsulated strains), H. parainfluenzae and H.aegyptius is normal flora of the upper respiratory tractInfections causing:1. Pyogenic meningitis2. Acute epiglottitis3. Cellulitis, middle ear infection,etc

4. conjuctivitis

5. MicroscopySmall, non-motile, Gram negative rods or coccobaciliLong thread-like form in old csf culture

6. Microscopic observation

7. Culture of H.influenzaeH.influenzae grows better in aerobically compare to anaerobicallyThe optimum temperature for growth 35 37oCThe are X and V factorBoth represent in blood agar and permit the culture to growH.influenzae and H.aegyptius need X and V factor, H. parainfluenzae need V factor and H.ducreyi need X factor

8. Biochemical testsNot usually used to identify hemophilus6 biovars of H.influenzae are recognized based on the indole, urease and ornithine decarboxylase (ODC) reactions of the diff strains

9. SerologyConsist of 1 f serotypesMostly causing meningitis belong to serogroup bMost strains that cause chronic bronchial disease are non-capsulatedAntimicrobial sensitivityResistant towards chloramphenicol, ampicilin, tetracycline, erythromycin and cotrimoxazoleH. ducreyi is sensitive to sulphonamidesAmpicillin resistant are common

10. Staphylococcus aureusStaphylococcus epidermidisStaphylococcus saprophyticus

11. Scientific classification Kingdom:Bacteria Phylum:Firmicutes Class:Bacilli Order:Bacillales Family:Staphylococcaceae Genus:Staphylococcus

12. Introduction Are microbial flora of the skin, upper respiratory tract and intestinal tract S.aureus usually cause abscesses, boils, conjuctivitis, pneumonia, septicemia, food poisoning and scalded skin syndrome S. epidermidis causing bactericaemia S. saprophyticus causing cystitis and acute urethritis

13. Laboratory diagnosisMicroscopy Non-motile Non capsulate Gram positive cocci Arrangement single or in pair Size 1 m diameter

14. Culture Grow well in aerobically and also in present of carbon dioxide Temperature between 10 420C, optimum temperature are between 35 - 370C

15. S.aureusProduce yellow to cream in blood and chocolate agar (heated agar)Occationally produce white 1-2 mm in diameter coloniesSome strain produce beta-hemolytic when grown aerobicallyColonies are slightly raised and easily emulsified on a slideNon- lactose fermenter in MacConkey agarMannitol salt agar is a useful differential and selective agar to identify S.aureus

16. On blood agar

17. S.epidermidisColonyis whiteNon hemolytic in blood agarS. saprophyticusMaybe white or yellowThere are non-hemolytic in BANot grow anaerobicallyNo growth in MacConkey agar

18. Biochemical reactionsS.aureusDNAse test will be positive for S.aureus but negative in other speciesCatalase test will be positive in all staphylococcus but negative in all streptococcusS. epidermidis and S. saprophyticusCoagulase negativeDNAse negativeCatalase positive

19. Antimicrobial sensitivityErythromycinClindamysinFucidinVancomycinMany strains of S.aureus are penicillin- resistantS.epidermidis are more resistant than S.aureus to antibioticsS. saprophyticus less resistant to antibiotics than S.aureus and S.epidermidis

20. Streptococcus pyogenesStreptococcus agalactiaeEnterococci

21. Lecture content Basic characteristics Classification Grouping of species Pathogenesis Lab diagnosis

22. Scientific classification Kingdom Bacteria Phylum - Firmicutes Class Bacili Order Lactobacillales Family Streptococcaceae Genus - Streptococcus Species 22 sp.

23. Basic characteristics Gram-positive Lactic acid bacteria group Spherical Grow in chain (twisted) or pairs Mostly are facultative anaerobes

24. Habitat Available in food such as Emmentaler (Swiss) cheese Normal flora of the mouth, skin, intestine and upper respiratory tract of humans

25. Classification

26. Alpha hemolysisSurrounded by an area of partial haemolysisGreen-brown colour (reduced haemoglobin)Consist of pneumococci and viridans group( alpha hemolytic and no Lancerfield antigens

27. PathogenicityS.pyogenes (Group A) Acute sore throat Scarlet fever Ear infections Peurperal sepsis Septicemia and occasionally endocarditis Skin infection such as celulitis and impetigo

28. Post-streptococcal diseaseFrom the implication of pathogenesis, it will leads to a serious stage of diseases: Acute glomerulonephritis Rheumatic fever

29. Enzymes & toxin productionBy S. pyogenes Streptolysin O Streptolysin S Streptokinase (cause fibrinolysis) Hyarulonidase DNAses DPNAse Erythrogenic toxin

30. Other speciesS.Agalactiae (Group B) Neonatal septicaemia, pneumoniae, meningitis Septic abortion and peurperal sepsisEnterococci (Group D) Urinary tract infection Infections of ulcer and wounds Endocarditis and meningitisViridans Endocarditis Dental caries Brain abscess

31. Microscopy Gram positive (purple) Non-motile Long chains Sometimes in pair

32. Culture Aerobically and anaerobically Temperature between (22 42o C) And enterococci are between (15 45o C) On BA, colony usually less than 1 mm in diameter Grey white or colourless Dry / shiny, irregular outline Most of the pathogenic streptococcus producing hemolysins

33. S. pyogenes Produce beta hemolysis Sensitive to bacitracin (cant rely 100% to it, coz not only group A sensitive to it) but for viridans it is resistance towards bacitracin A selective BA containing crystal violet was suggested Does not grow in MacConkey agar

34. S.agalactiae Optochin resistant but S.pneumoniae is sensitive Beta hemolysis in BA In serum starch agar, it produces orange pigment

35. Biochemical testSpecies Catalase Bacitracin Optochin Litmus CAMP MilkS.pyogenes - + - - -S. agalactiae - - - - +Enterococci - - - + -Viridans - - - - -streptococciPneumococci - - + - -