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STREPTOCOCCI
STREPTOCOCCI
Gram (+) spherical cells, arranged in chains or pairs Grayish pinpoint translucent to slightly opaque colonies Catalase (-), oxidase (-) Growth is enhanced by blood, serum or glucose incorporated in
STREPTOCOCCI
Behave-like facultative anaerobe, some species require increased CO2 for growth Aerotolerant anaerobe (peptostreptococci obligate anaerobe
STREPTOCOCCI
ALL streptococci EXCEPT the viridans group have a layer of C carbohydrate serological classification Young cultures are characterized by the presence of capsule Are commonly found as part of normal human flora.
STREPTOCOCCI
However, when these organism gain access to normally sterile sites they can cause life threatening infection. S. pyogenes and S. pneumoniae - notorious pathogens Aerococcus, Lactobacillus, Leuconostoc and Pediococcus resemble streptococci
STREPTOCOCCI
Colony variation a.Mucoid b.Matte or rough large amount of CHON c.Smooth/glossy little amount of CHON
CLASSIFICATION OF STREPTOCOCCI
1. PYOGENIC GROUP
Will not grow neither on 10C or 45C Produce pus; mostly -hemolytic Species: Streptococcus pyogenes
2. VIRIDANS GROUP
Will grow at 45C NOT part of the lancefield group Alpha hemolytic OR Non hemolytic Species: S. salivarius, S. mutans, S. mitis, S. sanguis
3. ENTEROCOCCUS
Will grow both at 10C and 45C Normal flora of human intestine Species: E. faecalis
4. LACTIC GROUP
Will grow on 10C Non hemolytic organism Often found in dairy products Species: S. lactis
CLASSIFICATION OF STREPTOCOCCI
1. ALPHA-HEMOLYTIC STREPTOCOCCI
Partial/incom plete hemolysis of RBC around colony Green discoloration around colony Species: S.
2. BETA-HEMOLYTIC STREPTOCOCCI
Complete hemolysis of RBCs around the colony Clear area/zone around colony Yellow discoloration Species: S. pyogenes, S. agalactiae
3. GAMMA-HEMOLYTIC STREPTOCOCCI
NO lysis of RBCs around colony/red cells immediately surrounding the colony are unaffected (no change) Species: E. faecalis (GROUP D)
C. LANCEFIELD BASED ON THE ANTIGENIC NATURE OF CELL WALL CARBOHYDRATES C POLYSACCHARIDE Rebecca Lancefield found out that C carbohydrate can be extracted from the streptococcal cell wall by placing the organism in dilute acid and heating for 10 min.
CLASSIFICATION OF STREPTOCOCCI
GROUP A STREPTOCOCCI
Pathogenic to man Not considered part of the normal flora Acquired thru contaminated droplets by cough or sneeze Species: S. pyogenes fever producing bacteria, flesh eating bacteria (involves deeper tissues and organs)
GROUP A STREPTOCOCCI
Principal virulence factor: Mprotein (type specific; antiphagocytic) Other virulence factor: a.Protein F mediates epithelial cell attachment b.Lipoteichoic acid bacterial adherence to the respiratory epithelium
3. SKIN INFECTIONS
CELLULITIS - is a diffuse, spreading infection of subcutaneous skin tissue characterized by a defined area of redness (erythema) and accumulation of fluid (edema)
3. SKIN INFECTIONS
ERYSIPELAS - is an acute infection and inflammation of the dermal layer of the skin characterized by painful reddish patches that enlarge and thicken with sharply defined edges - may also lead to necrotizing fasciitis and myositis galloping
3. SKIN INFECTIONS
RHEUMATIC FEVER ACUTE GLOMERULONEPHRITIS OR BRIGHTS DISEASE - inflammatory disease of the renal glomeruli; results from the deposition of antigen-antibody complexes.
DIAGNOSTIC TESTS
SULFAMETHOXAZOLETRIMETHOPRIM TEST
Positive: Any zone of inhibition Negative: No Zone of inhibition (Group A and Group B)
SXT
SXT
GROUP B STREPTOCOCCI
Part of the normal flora of female genital tract and lower GIT Nosocomially transmitted by unwashed hands of mother or healthcare personnel. Infection of fetuses and infants infection during passage through the colonized birth canal, and premature rupture of mothers membranes
GROUP B STREPTOCOCCI
On culture, it is grayish white mucoid w/ small zone of beta hemolysis Species: S. agalactiae Virulence factor: capsule (sialic acid signifiant component of the capsule) Enzymes: hemolysin, CAMP factor, neuraminidase, deoxyribonuclease, hyaluronidase and protease
DIAGNOSTIC TESTS
CAMP TEST
To differentiate S. agalactiae from other -hemolytic streptococci It uses -lysin producing strain of S. aureus or disk impregnated w/ lysin Result (+) arrowhead hemolysis near S. aureus growth /
GROUP C STREPTOCOCCI
Main source of streptokinase; animal pathogen Species: S. equi, S. equisimilis, S. dysagalactiae, S. zoopedimicus
GROUP D STREPTOCOCCI
Enterococci: E. faecalis, E. faecium, E. avium, E. gallinarium Non enterococci: S. bovis, S. equinus Enterococci are not highly pathogenic; cocci in pairs and long chains
DIAGNOSTIC TEST
LAP Test
Detects for the ability of the bacteria to hydrolyze Leucine-naphthylamide by enzyme leucine Positive: Development of a red aminopeptidase to -naphthylamide.
color within 1 minute after adding cinnamaldehyde reagent (S. Pneumoniae and S. pyogenes, Enterococcus, Pediococcus )
Negative: No color change or development of slight yellow color (Aerococcus and Leuconostoc)
NOTES
Enterococci (+) bile esculin, growth in 6.5% NaCl. PYR test and LAP test; vancomycin resistant Leuconostoc, Pediococcus, Globicatella, S. urinalis, Lactococcus (+) growth in bile esculin and 6.5% NaCl All enterococci isolated from human hydrolyze PYR
NOTES
Isolation of S. bovis from a blood culture may be the first indication that the patient has an occult tumor in the GI tract (GI cancer) 85% of Group B streptococci is (+) w/ growth in 6.5% NaCl but (-) in bile esculin test E. faecalis requires cyanocobalamin (B12) as growth factor.
GROUP F and G
Minute streptococci w/ wide zone of beta hemolysis Species: S. anginosus
VIRIDANS STREPTOCOCCI
Alpha hemolytic streptococci that lack the lancefield group antigens and do not fall on the criteria for S. pneumoniae, can be non-hemolytic. Oropharyngeal commensals Opporunistic pathogens of low virulence Glucans and dextrans enhance attachment to host cell surfaces
VIRIDANS STREPTOCOCCI
Laboratory tests: bile insoluble, optochin resistant, no growth in 6.5% NaCl, (-) inulin fermentation and PYR (-) Species: S. mutans, S. salivarius, S. mitis Infections: subacute endocarditis, dental carries (S. mutans)
ABIOTROPHIA
Pyridoxal-dependent or Vit B6 dependent and symbiotic streptococci Formerly known as nutritionally variant streptococci (NVS) Thiol compounds cystein, Vit B6 and pyridoxal This group of organism were found as satellites around an organism that produces pyridoxal E. coli, Klebsiella spp.,Enterobacter spp., yeasts
ABIOTROPHIA
They produce satellatism w/ Staphylococcus Opportunistic pathogens of low virulence Variable morphology gram (+) to gram (-) to gram variable; pleomorphic forms It causes 5-6% endocarditis
STREPTOCOCCUS SPP.
LABORATORY DIAGNOSIS
1. Gram Stain 2. Culture BAP, PEA, CNA, CAP For isolation of Group A streptococci from throat swab, BAP w/ SXT is used to suppress the growth of normal flora. To detect genital carriage of Group B streptococci during pregnancy, Todd Hewitt broth w/ antibiotics (gentamicin, nalidixic acid or colistin and nalidixic acis) is used to suppress vaginal flora.
Visualization of beta hemolysis is enhanced by anaerobic condition (BAP should be inoculated by stabbing the inoculating loop into the agar several times) CAP is incubated w/ 5-10% CO2 Abiotrophia will not grow on BAP or CAP unless pyridoxal (B6) is supplied. Streptococci has tendency to produce increase amount of lactic acid and other organic acid from fermentation of sugar resulting to acidic medium and eventually their growth is inhibited.
3. Susceptibility Test bacitracin disk test SXT test 4. CAMP and Hippurate test 5. Bile Esculin and Salt Tolerance Test 6. Serological Test a.Latex agglutination b.ASO titer test c.ELISA d.Co-agglutination
Streptococcus pneumoniae
Diplococcus/pneumococcus Gram (+) cocci in pairs, oval or lancet shape; facultatively anaerobe Causative agent of lobar pneumonia Considered part of the normal flora (25-50%) of the URT of preschool children Most common bacterial pneumonia in elderly as well as in patients w/ underlying disease.
Streptococcus pneumoniae
Most common cause of bacterial meningitis in adults Common isolate both as a pathogen and as a member of the normal respiratory tract. Principal virulence factor: antiphagocytic capsular polysaccharide type 3 strains Opsonization of the capsule renders the organism non virulent
Streptococcus pneumoniae
The capsule is antigenic and can be identified w/ appropriate antiserum; it is composed of hyaluronic acid. Alpha hemolytic colonies, mucoid, flattened with depressed center. dome-shaped, glistening, wet, mucoid young colonies coin w/ a raise rim old colonies (due to autolytic
Streptococcus pneumoniae
Dimple-shaped/donut-shaped colonies Enzymes: hemolysin, immunoglobulin A protease, neuraminidase and hyaluronidase
INFECTIONS/DISEASES PRODUCED
1. Pneumonia - pneumonia due to S. pneumoniae is not usually a primary infection but rather a result of disturbance of the normal defense barriers. 2. Meningitis 3. Otitis media 4. Bacteremia 5. Endocarditis 6. Peritonitis
Differential Test 1. Optochin Susceptibility/P disk Test Presumptive test Ethylhydroxycuprein hydrochloride BAP (sensitivity media) 2. Bile Solubility Confirmatory test Differentiates pneumococcus from viridans streptococci Sodium desoxycholate When a heavy suspension of pneumococcus is added to bile salt, the cloudiness of the broth clears after 3 hrs incubation.
Result
> Or = 14mm
(+) soluble
Differential Test 3. Neufeld Quellung Reaction The antipnemococcal serum is mixed w/ sputum, CSF and other sources along w/ methylene blue Examined under OIO
Result
SWELLING CAPSULE