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state of the host 1. The organisms compete with pathogenic organisms for potential attachment sites. 2. These organisms often produce substances (toxins or acids) which are bactericidal. OBSTACLES OF MICROORGANISMS • The microorganism must avoid being caught up in the mucus layers of the upper respiratory tract, being transported to the back of the throat, and eventually being swallowed. • If the invader has deposited in the lower respiratory tract or lung, it must either avoid phagocytosis, or be able to survive and multiply in the phagocytic cell. • Airborne disease transmission • inhale 10,000 to 20,000 L of air per day • main route of transmission of pathogens • droplets become airborne • normal breathing • coughing and sneezing STREPTOCOCCUS PNEUMONIAE General Features - gram (+), non motile, encapsulated cocci - lancet shaped, in pairs - fastidious - common cause of respiratory infections Epidemiology - found nasopharynx - sensitive to environmental agents - endogenous – carriers due to impaired immune system - exogenous – droplet transmission Predisposition to pneumococcal infection -defective Ab formation -insufficient numbers of PMNs -day-cares, military, prisons, shelters -chronic respiratory disease -infancy and aging -diabetes, alcoholism, liver disease Virulence Factors 1. Capsule - antiphagocytic and antigenic - 85 capsular serotypes 2. Autolysin - peptidoglycan hydrolase - responsible for release of intracellular virulence factor 3. Pneumolysin - attacks mammalian cells Clinical Significance • Acute bacterial pneumonia • otitis media • bacteremia / sepsis • meningitis
Laboratory diagnosis of Strep. pneumoniae
Culture in BAP with 5% CO2 α hemolytic
direct Gram staining gram (+) diplococci
Sputum + small amount of broth in 1-2 ml syringe
“ candle jar: • Optochin test • Bile solubility test sensitive
Type organism from Peritoneal washing
Treatment - penicillin resistance due to alteration of PBP - 3rd generation Cephalosporins - Vancomycin Vaccine Pneumococcal vaccine 23 different serotypes account for 90% of invasive strains protection wanes with time and age Indications for vaccine advanced age myeloma splenectomy alcoholism HIV / AIDs diabetes lymphoma PREVNAR / SYNFLORIX - conjugate vaccine - indicated for use in infants < 2 years of age STREPTOCOCCUS PYOGENES (STREP THROAT) - Streptococcus pyogenes – most virulent - can invade intact skin or mucous membrane - low inoculum cause infection - nasopharyngeal carriage common Characteristics - gram (+), nonmotile cocci - occur as long chains Virulence Factors 1. Capsule – hyaluronic acid 2. Cell wall a. fimbriae - contains M protein - N – terminal end has antiphagocytic activity - M proteins highly variable - lipoteichoic acid – adherence b. C – carbohydrate - rhamnose and N-acetylglucosamine c. Protein F ( fibronectin – binding protein ) - mediates attachment 3. Extracellular products a. Streptococcal pyrogenic exotoxins (SPE) - SPE A and C - carried by bacterophage - SPE B- cysteine protease b. Cytolytic toxins and other exoenzymes
synthesis of iron containing respiratory enzymes 2. Impetigo 4. aphrophilus – X factor • H. hypotension. pyogenes infection .small.damages cells resulting in release of lysosomal enzymes and degranulation . lesion samples. prevents opsonization & phagocytosis Damages vascular epithelium and blood brain barrier Degrades IgA Disrupts the beating of cilia on epithelial cells of respiratory tract May induce hypotension. polyribose ribitol phosphate (PRP) . influenzae . parainfluenzae – V factor • H. sputum.1889 – 1892 . fever.strongly antigenic Streptolysin S . Puerperal sepsis .sensitive: Bacitracin . Invasive Grp A Streptococcal Disease . X factor – haemin .both • H.Serologic test: ASO test.1933 – Smith.may colonize the conjunctiva and genital tract . Andrew and Lardlaw.Grp A C-carbohydrate . V factor – NAD/NADP .deep local invasion with or without necrosis .i.1883 first documented by Robert Koch . shock and other systemic symptoms Peptidoglycan IgA proteases Exotoxin-like factor Lipopolysaccharide .etiology of influenza is a virus General Features -gram (-).humans only natural host . Anti DnaseB titer Treatment drainage and debridement Antibiotics: Penicillin Macrolides HAEMOPHILUS INFLUENZAE (CHILDHOOD PNEUMONIA) Historical Background . small uniform coccobacillus -aerobic or facultative anaerobic .purulent inflammation of posterior oropharynx and tonsillar area 2.Pfeiffer noted small bacilli in sputum .e.normal flora of upper respiratory tract nonencapsulated strain in 25 – 80% capsulated strain in 5 – 10% . Acute Glomerulonephritis Laboratory Diagnosis .2-3 weeks after strep throat 8.most common type of S. Acute Rheumatic Fever . ducreyi – X factor Pathogenesis .Necrotizing Fascitis / Myositis .lyses erythrocytes.catalyzes conversion of plasminogen to plasmin Streptodornases C5a peptidases Hyaluronidase Clinical Significance 1. blood or CSF .Streptolysin O . pus.hexosamine Type f .galactosamine type b capsule -repeated disaccharide units of ribose and ribitol phosphate . leukocytes and platelets Streptokinase . opalacent colonies . Cellulitis 3. multiple organ injury and rash 7. Acute pharyngitis or pharyngotonsillitis .encapsulated -demonstrated by capsule stains and Quellung reaction 6 distinct antigenic type Type a – glucose Type d .immunization reduced incidence Factors Associated with Virulence Factor Type b capsule Comments Major virulence factor Associated with invasive disease.oxidation – reduction processes • H.due to exogenous and endogenous transmission 6.ß hemolytic .toxic shock-like syndrome.associated with most invasive disease Growth Factors 1.1893 – published a report on Hemophilus influenzae as cause for influenza .precipitin reaction . Erysipelas 5.hexose Type b – ribose Type c – galactose Type e .influenza pandemic .throat swab.
Non Invasive Infection 1.supplemented with X factor .cause irreversible shock and cardiovascular collapse .synthesized by other organisms.incubate in aerobic atmosphere enriched with 5 – 10% carbon dioxide Chocolate Agar -heated at 80oC x 15 minutes .produces toxic products .gram (-) coccobacilli . Septic arthritis 5.associated with pharyngitis or septic arthritis or localized disease B. i.Lyses RBC to release factor X and V .80 – 90% of non.paroxysms of cough e.fastidious . epiglottitis . acute exacerbations of chronic airway obstructing disease Laboratory Diagnosis Direct examination Gram stain: small gram (-) coccobacilli. sinusitis 3.immune sibling exposed at home .Presumptive identification Culture General Considerations . moist.e.agglutination property – important for identification .inhibits phagocytosis and NK cell function d.facilitates adhesion b. Lipopolysaccharide (LPS) .latex agglutination test.counterimmuno electophoresis .otitis media 2. Dermonecrotic (heat-labile) toxin .< 50% of non-immune child contacts at school Virulence Factor Polysaccharide capsule .(+) capsule in virulent strain .small.very strong vaso-constrictor and causes ischemia and extravasation of leukocytes .Respiratory droplet Degree of contact . aureus on BAP Immunologic Assay .for optimal yield: transport immediately and inoculate on appropriate media without delay . bacterial meningitis 4.accumulation of large amounts of cAMP which leads to increased mucus secretion and interferes with many cellular functions c.cause tissue necrosis and airway obstruction .causes necrosis of the tracheal tissue f.inhibits many leukocyte functions. Cellulitis 6.life threatening 2.Add drop of S. Tracheal cytotoxin .binds to ciliated epithelial cells -. Staphylococcus Presumptive Diagnosis. phagocytosis and impairs NK cell killing . filamentous hemagglutinin .rapid detection of free. Invasive Infection 1. butyrous consistency Mode of transmission . Adenylate cyclase .strict aerobe .Inactivates enzymes that could destroy factor V .prevents the deactivation of adenylate cyclase .major cause of pertussis (abnormal cough) . greyish-white convex. Pneumonia 3.isolates grow only in presence of X and V factors Satellite Phenomenon.chocolate agar .Sensitized particles with antibody against specific antigen Definitive Diagnosis – use biochemical test – molecular techniques BORDETELLA PERTUSSIS (WHOOPING COUGH) General Features .increase histamine release and enhance inflammatory response .contributes to bacterial binding to ciliated epithelial cells .peak at 2 – 4 years old . including chemotaxis.paralyses cilia .protects from bactericidal processes Toxin production a.optimal growth at 35 – 36 o C -typical colonies: shiny.Fimbriae Adhesins Assist in adherence Clinical Significance A.inactivates substances that inhibit growth .viability decrease with time and in 40 C .cellulitis of epiglottis and surrounding tissues . Bacteremia . soluble form of type b capsule . Pertussis toxin . transparent colonies * V factor – present in other biologic fluids .
Modified Tinsdale or cystiene tellurite media . heat and tenderness • laryngeal and bronchial diphtheria Laboratory Diagnosis Schick test . fatigue. etc.lethal dose: 0.raised.young children 0 1 2 3 4 5 6 7 8 9 10 Weeks Diagnosis . dew drops or mercury droplets .Cough plates Bordet – Gengou agar Where is the best place to swab? perinasal area • fewer commensals • tip directed downwards and toward the midline ( 5 cm ) • Collect specimen on several successive days for better isolation ( 80%) Bacterial Culture • highest specificity • enable to serotype and genotype • organisms can be recovered up to 3 months of illness • supplementation with antibiotics – penicillin ( 0.activate a variety of inflammatory mediators ( TNF. IL1.Susceptible localized erythema maximum size and intensity ( 1-3 cm) in 2-4 days No neutralizing toxin Specimen collection . IL6. non capsulated • Neisser or Albert stains – beaded appearance Pathogenesis • prevalent in Third World countries • confined to humans and horses • direct spread from nasopharyngeal secretions • peak incidence .nasopharyngeal washing / swab Dacron or Calcium alginate swab do not use Rayon or cotton –(+) fatty acid . fever Cough.Symptoms are characteristic Laboratory diagnosis . stabilized toxin .1 ug/kg . fever and sore throat • thick nasal discharge • tonsillar infection – thick .use holding media trycase agar with 5% sheep blood cystine-tellurite modified Tinsdale Loeffler’s slant blood or serum tellurite media Tellurite_ inhibit growth of normal flora . transparent borders .) and generate complement activation products Clinical Presentation of Bordetella pertussis disease Incu Catarrhal Paroxysmal batn Phase Phase Colds.throat and nasopharyngeal areas .do not force swab .15 mg/l) – cephalexin (30mg/l) • Colonies ..2 fragments Fragment A – inhibits protein synthesis Fragment B required for transport of A Signs and Symptoms • malaise.Obtain nasopharyngeal aspirate and primary culture on Bordet-Gengou medium (potato-glycerol-blood agar) .intradermal injection of diluted. adherent. smooth. prostaglandins.small zone of hemolysis . malaise. vomiting leukocytosis Convalescence Intensity of symptoms • V shaped or palisades • ‘chinese lettering’ • pleomorphic • gram (+) • aerobic and facultative anaerobic • grows best at 37oC on blood containing medium • non motile.bisegmented pearl.heat labile .1 mm . non sporeforming.appear at 48 – 72 hours Treatment: • drug of choice: erythromycin • may also be given to household contacts and nonvaccinated infants CORYNEBACTERIUM DIPHTHERIAE (DIPHTHERIA) General Features Diphtheria toxin . green pseudomembrane • ‘bull neck’ – edema.does not grow on common laboratory media -oxidase positive but urease negative .grows as small transparent hemolytic colonies Specimen collection .screening test .
shape and color of colonies used to differentiate 3 biotypes gravis – large nonhemolytic colonies intermidius – small colonies mitis – larger hemolytic colonies biochemical tests . jeikeium C. Minitek and Biolog hemolysis Nitrate reduction Organism catalase maltose sucrose V + V glucose urease C.Loeffler’s slant – quick growth – provide characteristic morphology Size. sugar fermentation and gelatin within 5 days Others .enzymatic results in 24 hours . urealyticum A. hemolyticum MYCOBACTERIUM TUBERCULOSIS Characteristics • Obligate aerobe • Intracellular organisms • Slow growing takes 12-20 hours to divide • Bacillus • Weakly gram positive cell wall • Shows beaded appearance.urease. minutissimum + + + + + + + + - V + + + + + + + + - + V + + - + + + + + + + + + + + + V + + C. xerosis C.most useful .20 bichemical test in a strip .Vitek.to differentiate from other coryneforms Commercial Kits API Coryne System . pseudodiphtheri C.slow growth – greyish to black colonies . diptheriae C..glycogen metaphosphate granules xylose . pseudoTB C. ulcerans C.
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