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Chapter 8 Coccus Pathogenic cocci • pyogenic coccus • according their staining colour – G(+) : Staphylococcus, Streptococcus, Pneumococcus – G(-) coccus : – Meningcoccus, Gonococcus The Staphylococci Staphylococcus Morphology Morphology : Shape : Gram staining : Special structure : Distribution : Kinds: S. aureus Staphylococcus Culture 1.Liquid medium: Turbidity growth 2.solid medium: turbidity growth Oil colony(gold、white、lemon) S.aureus : Blood agar plate (βhemolysis) Staphylococcus Biochemical reaction (-) (+) (-) (+) catalase test mannitol test Staphylococcus Antigenic structure ★staphylococcal protein A, SPA Capsule polysaccharide SPA Staphylococcus resistance – More resistance than other non-sporeforming bacteria – Variably sensitive to many antimicrobial drugs – Drug-resistant strains increase, and become the most common hospital pathogen control drugtreated Main characteristics of three species of staphylococci property S aureus S epidermids S saprophyticus Pigments of colony Gooden yellow White White or gray Coagulase + - - Α-hemeolysin + - - SPA + - - Carbohydrate + + - Phage typing + - - pathogenicity + +/- - fermention Staphylococcus Toxin and enzymes 1. coagulase 2. heat-stable nuclease 3. staphylolysin 4. leukocidin 5. enterotoxin 6. exfoliative toxin, exfoliatin 7. toxic shock syndrome toxin 1,TSST-1 8.Other enzymes Staphylococcus hyaluronidase hyaluronic acid Pathogenesis of staphylococcal infections Staphylococcus pyogenic inflammation food poisoning scalded skin syndrome toxic shock syndrome, TSS Pseudomembrane colitis,PMC furuncle impetigo Folliculitis carbuncle Microbiologic diagnostic test specimens pus, blood, tracheal aspirate, et.al smears G+ Staphylococci isolated culture Growth phenomenon Biochemical reaction pigment,hemolysis Catalase test Coagulase test Susceptibility test The Streptococcus I. Biological characteristics Streptococcus Morphology Morphology : Shape: Special structure: Gram staining: (1).Hemolytic activity: (i) -hemolytic streptococcus *Incomplete hemolysis, green coloration of the medium surrounding the colony. *Opportunistic pathogens – subacute bacterial endocarditis (SBE). (ii) -hemolytic streptococcus (or pyogenic streptococcus) Complete hemolysis, major human pathogens (iii) -streptococcus No hemolysis, no pathogenicity. Staphylococcus Culture 1.Liquid medium: 2.Blood agar plate : Long chain and deposit S. pyogenic Antigenic structure (group-specific antigen). 19 groups Group A streptococci are main human pathogens Polysaccharide Ag M protein type-specific antigen M protein: *presents in cell wall (group A) *Anti-phagocytosis *adhere to epithelial cells *clump platelet and leukocyte *heat stable; acid stabl S. pyogenic resistance *heat labile: 60℃,30 min *antibiotics sensitivity: penicillin G ,etc. II. pathogenicity (1).Invasiveness (i).surface structure *LTA(lipoteichoic acid): adhere to sensitive cell (epithelial cell; platelet; RBC; WBC; lymphocyte; mucous membranes) F protein * M-protein : anti-phagocytotic M-Ag Ab hypersensitivity (glomerulonephritis) Common antigen--- cross-reacts heart myosin autoimmunity------- rheumatic fever Toxins and enzymes Hyaluronidase (spreading factor): – Pyrogenic Exotoxins (ErythrogenicToxin) Streptokinase (SK ): Streptodornase (SD): – streptolysin,SL • S. pyogenic Pathogenesis & Clinical Findings 2. Diseases of streptococcal infection (1). local purulent infections: (2). systemic infection : Erysipelas Scaret fever Tonsillitis (3). Hpersensitivity response : (i) acute glomerulonephritis ( group A) (many types of group A streptococci) mechanism: *immune complex (deposition) glomerular basement membrane type III hypersensitivity *common Ag cross-reacts glomerular basement type II hypersensitivity (ii) Rheumatic fever (many types of group A streptococci) mechanism: *immune complex (deposition) heart, joints type III hypersensitivity *common Ag cross-reacts heart type II hypersensitivity S. pyogenic Diagnostic Laboratory Tests • Specimens – A throat swab, pus, or blood • Smears – Smears from pus often show single cocci or pairs rather than definite chains. • Culture • ASO test: ASO titer > 1: 400 units, help to diagnose rheumatic fever. Streptococcus *Treat the pharyngitis and tonsillitis in time, avoid streptococcal hypersensitive disease. *Antibiotics and chemical agents: penicillin G for the first choice S. pneumoniae S.pneumoniae Morphology Morphology : Shape: Special structure: Gram staining: S.pneumoniae Culture 1.Blood agar plate : 2.Liquid medium: S.pneumoniae Biochemical reaction Not optochin sensitive • α-hemolytic streptococcus and • S.pneumoniae Bile bacteriolysis test Optochin sensitive test optochin sensitive S.pneumoniae Capsule prominent virulent strains S.pneumoniae R型 S型 ? Pathology • pneumococcal pneumonia ---pneumonia Diagnostic Laboratory Tests • specimen – Blood cultures, sputum, pleural fluid, and spinal fluid • Stained Smears: – A Gram-stained film of rusty-red sputum shows typical organisms, many polymorphonuclear neutrophils, and many red cells. • Capsule Swelling Tests – identification of pneumococci and possible typing • Culture--- Sputum cultured on blood agar Therapy – most strains susceptible to penicillin a large daily dose of penicillin – resistance is common Drug sensitivity testing is therefore essential – Polyvalent vaccines that contain capsular antigens to preventing recurrences Neisseria • Gram negative • diplococci • oxidase positive • Culture: chocolate agar * heated blood Gram stain of pure culture exudate Using the Gram stain in patient specimens, the organisms are most often observed in polymorphonuclear leukocytes N. meningitidis Main Pathogenic factor: LPS Capsule have Plasmid rarely have Caused disease meningitis N. gonorrhoeae PILI not have have genital infection prostatitis Urethritis cervicitis Endometritis Purulent conjunctivitis Infection in newborns during vaginal delivery blindness Prevention and treatment *penicillin----*silver nitrate----conjunctivitis 1. Lancefield Grouping of Streptococci (Group A, B, etc.) is based on: √A.Cell wall carbohydrate B.M protein C.LPS D.Peptidoglycan E.DNase production 2. Vaccination against the Streptococcus pneumoniae capsule is protective against pneumonia because: A.There is only a single serotype of pneumococcal capsular antigen. √B.Anti-capsular antibody opsonizes the organisms in the lung so they can be phagocytosed. C.It stimulates cell-mediated immunity so that the organisms are killed in the lung. D.It prevents the pneumococci from colonizing the throat and then getting into the lung. E.It inhibits the production of alpha hemolysis. 3. Acute Rheumatic Fever: A.Usually appears 3 weeks after primary skin infection with Group A streptococci. the result of a cross-reactive immune response to √B.Is streptococcal antigens. C.Is manifested only as inflammation of the heart muscle. D.Will not occur if symptoms of primary streptococcal disease disappear within 3 days. E.NONE OF THE ABOVE. 4. You treat a patient with pharyngitis caused by Group A streptococci with amoxicillin. The pharyngitis resolves, but now the patient experiences a vaginal yeast infection. Which is the most likely explanation for this phenomenon? A. The pharyngitis was really oral thrush and the yeast spread through the alimentary and genital tracts. B. The Streptococcus pyogenes bacteria were keeping the yeast in check. C. The antibiotic induced a hypersensitivity reaction. √D. The normal flora that controlled the yeast was inhibited by the antibiotic. E. None of the above are likely. 5.To differentiate the viridans Streptococci from Streptococcus pneumoniae, which of the following tests is done? A. A co-agglutination test with Staphylococcus aureus and antibody to the pneumococcal capsule B. A test that examines resistance to bacitracin C. A coagulase test D. √ A test that examines sensitivity to optochin. E. A test that examines resistance to novobiocin. 6 .To determine if a patient with a newly diagnosed heart murmur had a recent Group A Streptococcal infection that could have caused rheumatic fever, which is the most effective test to be done? A. Throat swab and culture, looking for beta hemolytic colonies. √B. An M protein antibody test. C. An anti-DNAase B test. D. Examine the patient for a red, sandpaper-textured rash. E. Blood culture. 7.Which of the following can produce autoenzyme: A.Bacillus proteus B.Bacillus typhi murium C.Vibrio cholerae D.Meningococcus √ E.Clostridum tetani 8.The bacteria which can produce SPA is √A.Staphylococcus aureus B.Meningococcus C.Gonococcus D.aeruginosus Bacillus E.none of above 9.The test which can differentiate S.aureus from other species is √A. coagulase-test B.Catalase test C.Lactose ferment test D. mannitol test E.none of above 10.Which of the following substances are produced by S.aureus except A. coagulase B.staphylolysin C. leukocidin D. exfoliative toxin √E.streptokinase