What are the two medically important genus within the family Micrococcaceae?
Micrococcus
Staphylococcus
what is the species under the genus Micrococcus?
M. luteus
what are the three species under the genus staphylococcus?
S. aureus
S. epidermidis
S. saprophyticus
what are the six biochemical tests for the family Micrococcaceae?
Catalase
O-F Glucose
Coagulase
Novobiacin
Mannitol Salt Agar
DNAse Test
this enzyme catalyzes the liberation of water and oxygen from hydrogen peroxide, a metabolic end-product toxic to bacteria.
Catalase
this is the metabolic pathway used by microorganism during utilazation of a substrate, which results in an acid-by-product.
O-F Glucose
this "cluping factor" is a cell-associated substance that binds plasma fibrinogen causing agglutination.
coagulase
if coagulase is negative on the slide test it has to be confirmed on what?
tube
which coagulase test is more accurate, slide or tube?
tube
what is the dosage of novobiacin used for GPC testing?
5 mcg
novobiacin is an ___________ determination to check the microorganism's sensitivity to Novobiacin.
antimicrobial
Mannitol salt agar with 7.5% NaCl is used as a ____________ test.
tolerance
this is an enzyme that depolymerizes DNA resulting in its decomposition from macromolecule to a simpler form.
DNAse test
DNAse test is ude to ID what?
S. aureus
this is a family of gram positive cocci that is catalase positive.
Micrococcaceae
Micrococcaceae are what four shapes?
Clusters
pairs
chains
tetrads
Organisms in the family micrococcaceae have a ____________ appearance.
butyraceous
organisms in the Micrococcaceae family are what kind of oxygen users?
aerobic
faculatitive anaerobic
This term is applied to the Micrococcaceae family becuase all strains grow in the presence fo 5% NaCl.
Halophilic
Many organisms in the family Micrococcaceae will grow in _______ NaCl.
10-15%
this species in the micrococcaceae family is an obligate aerobe that lives in soil, dust, and water. Cells are typically arranged in tetrads and/or sarcinae.
Micrococcus luteus
M. luteus is rarely __________ except in compromised hosts.
pathogenic
what are the three conditions that M. luteus can be seen in the human body?
urinary tract infections
Endocarditis
Wound infections
this is a slow grower in the micrococcaceae family with a bright lemon-yellow, butyraceous, and convex appearance.
M. luteus
what is this organism?
-O-F Glucose: oxidizer
-Catalase: +
M. luteus
this organism in the micrococcaceae family is normal flora in the skin, nasopharynx, anterior nares, and perianal area, yet is clinically significant in wound infections, boils, carbuncles, abscesses, pustules, septicemia, TSS, food poisoning and secondary invasions.
Staphylococcus aureus
what are the six extracellular enzymes found in S. aureus?
coagulase
Hyaluronidase
Protease
Lipase
Penicillinase
Protein A
these four toxins found in S. aureus cause small veins to constict, post ischemic necrosis, and destroys RBCs, neutrophils, macrophages, plt.
Alpha, beta, gamma, and delta toxin
what is the toxin found in S. aureus that kills leukocytes?
leukocidin
this is the toxin that cuases toxic shock syndrom which is found in S. aureus.
TSST 1
this toxins in S. aureus are specific for intestinal cells.
enterotoxins
this toxins in S. aureus are responsible for skin sloughing and scaly skin.
exfoliatin
the colonial morphology for this Micrococcaceae organism on a BAP is:
-white or golden yellow
-1-3mm diameter
-non-hemolytic, or B/hemolytic
-circular, entire, convex
-butyraceous
S. Aureus
what is this organism?
Coagulate: +
Novobiacin: -
Mannitol fermentation: + (usually)
DNase: + (usually)
S. aureus
this organism in the micrococcaceae family is normal flora on the skin and mucous membrane, but clinically significant in wound infections, UTIs, SBE, artificial shunts and valve infections, and nosocomial bacteremia.
Staphylococcus epidermidis
this organism in the Micrococcaceae family is occasionally beta hemolytic, with a yellow or white appearance on a BAP, otherwise very similar to S. aureus.
S. epidermidis
what is this organism?
-Coagulase: =
-novobiacin (5mcg): S
-Mannitol fermentation: =
-DNase: =
S. epidermidis
this organism in the Micrococcaceae family is normal flora on the skin and GU tract, but clinically significant in UTIs in young sexually active females.
Staphylococcus saprophyticus
which organism has a similar colonial morphology on a BAP to S. epidermidis?
S. saprophyticus
what is this organism?
-Coagulase: =
-Novobiacin (5mcg): R
-Mannitol: + (usually)
-DNase: =
O-F glucose: oxidizer
S. saprophyticus
what is the reagent used in the catalase slide test?
3% hydrogen peroxide
when performing a catalase slide test how many drops of 3% H2O2 should be used for one colony?
one drop
on the catalase slide test, rapid furious and sustained bubbling indicates what reaction?
Positive
on the catalase slide test, no bubbling indicates what reaction?
negative
what two things could cause a false positve reaction on the catalase slide test?
iron-containing loops
picked up erythrocytes from BAP
what are the control organisms for Catalase test?
(+) Staphylococcus aureus
(=) Streptococcus pyogenes
what is the media used for O-F glucose test?
two O-F glucose tubes
how many times are the O-F glucose tubes stabbed with bacteria?
5 times
how much mineral oil should be used to cover the closed O-F glucose tube?
2ml
what temperature should O-F glucose tubes be incubated at?
35oC
in the O-F glucose test, yellow color on top portion of exposed tube indicates what reaction?
oxidizer
in the O-F glucose test, yellow color on both tubes indicates what reaction?
fermenter
in the O-F glucose test, no color change indicates what reaction?
non-utilizer
what are the QC organisms for O-F glucose and their results?
Fermentative: E. coli
Oxidative: A. anitratus
Negative: A. lwoffi
what is the reagent for the coagulase test?
Rabbit or Human plasma (EDTA)
why is rabbit plasma preferred in the coagulase test?
it clots more rapidly
what is the principle for the coagulase test?
prothrombin-like activity
what are the two forms in the coagulase test?
bound coagulase (clumping factor)
free coagulase
this occurs in a coagulase test when they are suspended in plasma on a slide test, (bound to the bacterial cell wall and reacts directly with fibrinogen.
bound coagulase
this is when coagulase is released by bacterial cells into culture filtrates in tube test, similar to that when thrombin is added.
free coagulase
free coagulase is secreted ____________.
extra-cellularly
free coagulase is released by bacterial cells into culture filtrates in _____ test.
tube
the coagulase slide test is also known as what?
bound coagulase
clumping factor
in the coagulase slide test, granulasr precipitate indicates what reaction?
positive
in the coagulase slide test, absence of clumping indicates what result?
negative
int he coagulase tube test complete or partial clot indicates what?
positve
in the coagulase tube test, absence of clot or gel indicates what result?
negative
strong coagulase positive organisms may produce a clot in __ to ___ hours?
1-4
delayed positive many occur only after ___ hours in coagulase test.
18
in coagulase this may dissolve the clot soon after it is formed.
fibrinolysin
clumped colonies can cause a false _________ in slide test coagulase.
positives
what are the controls for coagulase test?
(+) Staphylococcus aureus
(=) Staphylococcus epidermidis
tube coagulase tests should be incubated at 35oC for how long?
four hours
what are the intervals for checking a tube coagulase test?
30 min
1 hr
2 hr
3 hr
4 hr
what two medias can a novobiocin disk test be performed on?
BAP
MH
what is the method used for disk tests?
disk diffusion method
what are the quality controls for novobiocin tests?
resistant: S. saprophyticus
Susceptible: other staph spp.
what is considered a resistant result for novobiocin?
<12mm
what result is considered susceptible for novobiocin?
> 12mm
what is the pH indicator on the Mannitol salt agar?
phenol red
this determines microorganism's ability to ferment mannitol.
mannitol salt agar test
on an MSA, good growth of _______ colonieks indicates that organism ferment mannitol
yellow
what are the quality control organisms for mannitol salt agar?
S. aureus/saprophyticus
S. epidermidis
what is another name for microdase test?
modified oxidase
the microdase test is used to confirm what organism?
M. luteus
the microdase test should be incubated at room temp for how long?
2 minutes
pruple to dark blue color on a microdase test indicates what reaction?
positive
yellow to colorless on a microdase test indicates what result?
negative
what are the QC organisms for the microdase test?
Positive: M. luteus
Negative: S. epidermidis
what are the five organisms in the family streptococcaceae?
S. pyogenes
S. agalactiae
S. pneumoniae
S. viridans
E. faecalis
This organism is also known as Beta strep grp. A.
S. pyogenes
this organism is also known as beta or gamma strep grp. B.
S. agalactiae
this organism is also known as grp D or non enterococus.
E. faecalis
what are the eight biochemical test used on the family streptococcaceae?
PYR
Bacitracin (A)
Optochin (P)
CAMP
Hippurate hydrolysis
Bile solubility
Esculin hydrolysis
Salt tolerance test
this test is used to seperates S. pyogenes (+) and enterococcus (+) from other strep.
PYR
this test is also called the A disk.
bacitracin
this test is used to ID S. pneumoniae.
optochin
this test is also called the P disk
optochin
these two tests are used to ID S. agalactiae.
CAMP
Hippurate hydrolysis
this test is used on alpha hemolytic streps.
bile solubility
this test is used in the ID or group D strep.
esculin hydrolysis
this test is used to ID the subgroup of group D.
salt tolerance test
what is the genus for streptococcaceae?
streptococcus
what fastidious family of GPC requires blood based media and lactic acid from carbs
streptococcaceae
this family of GPC forms chains in liquid media.
streptococcaceae
streptococcaceae family is catalase ________, facultative anaerobics.
negative
the primary infections of this organism include:
scarlet fever
erysipelas
septicemia/bacteremia
meningitis
streptococcus pyogenes
This determinant of pathogenicity in S. pyogense is involved in attachemnt.
Protein F
this determinant of pahtogenicity in S. pyogenes is anti-phagocytic.
protein M
this determinant of pahtogenicity in S. pyogenes is invovled in fibrinolysis.
streptokinase
this determinant of pahtogenicity in S. pyogenes is oxygen labile, heat stable.
streptolysin-O
this determinant of pahtogenicity in S. pyogenes is oxygen stable, heat labile.
steptolysin-S
S. pyogenes may have autoAb that attack ________ tissue.
heart
S. pyogenes may have an Ab-Ag complex that builds up in the ________.
kidneys
S. pyogenes are pinpoint, circular, convex, and translucent organisms that are ______ hemolytic.
beta
what is this organism.
-Catalase: =
-Beta hemolytic
-PYR: (+)
-Bacitracin: (s)
-Hipp hydro: =
-CAMP (varibale)
S. pyogenes
This strep is normal flora in the upeer respiratory tract, lower GI, and female GU tract, but clinically significant in wound infections, neonatal meningitis and sepsis, and puerperal sepsis.
Streptococcus agalactiae
this GPC is similar to S. pyogenes but has a smaller zone of hemolysis and some strains are gamma.
S. agalactiae
What is this organism?
-Catalase: =
-Beta or gamma hemolytic
-Bacitracin: R
-Hipp hydro: +
-CAMP: +
S. agalactiae
what groups of strep are considered other/misc. step?
C, F, & G beta hemolytic
what group C strep clinically significant for?
acute pharyngitis
what is group G strep clinically significant for?
malignancies
these are three strep groups which are very similar, grayish white, glistening, with variable hemolysis.
Grp C, F, & G beta hemolytic
ID of these strep groups is unimportant but must be differentiated from grp A by PYR.
group C, F, & G beta hemolytic
this strep was formerly genus diplococcus.
S. pneumoniae
this is the major cause of bacterial pneumonia.
S. pneumoniae
what are two synonyms for S. pneumoniae?
pneumococcus
Diplococcus
What is the most common clinical significance for S. pneumoniae?
Meningitis
S. pneumoniae has a ______ which enhances virulence.
capsule
This frequently encapsulated GPC is lancet shaped and stimulated by CO2.
S. pneumoniae
what is the colonial morphology for encapsulated S. pneumoniae on a BAP?
small
mucoid
alpha hemolytic
oil/water droplets
what is the colonial morphology for NON-encapsulated S. pneumoniae on a BAP?
small
glistening
alpha hemolytic
convex (young)
umbonate (old)
how can young and old S. pneumoniae be deferentiated on a BAP?
young-convex
old-umbonate
what is this organism?
Catalase:=
alpha hemolytic
optochin (P): S
bile solubility: +
S. pneumoniae
this group of strep is not usually speciated and is clinically significant in subacute bacterial endocarditis.
Viridans group
this alpha or gamma hemolytic GPC in chains is usually has a pinpoint, smooth or matte appearance on a BAP.
viridans strep
What is this organism?
Catalase: =
optochin: R
bile: =
growth on 6.5% NaCl: =
hippurate hydrolysis: =
Viridans group
this organism is normal fora of birds, animals, and humans, it can be found on large bowel and female GU tract. it is clinically significant in peritonitis.
enterococcus
this organism is small less opaque and can be glossy-white. may produce beta, alpha, or gamma hemolysis on a BAP.
enterococcus
what is this organism?
Catalase: =
Bacitracin: R
Hipp Hydro: =
CAMP: =
optichin: R
Bile: =
BEM: +
6.5% NaCl: +
Enterococcus
this classification scheme is based on temperature group growth characterisitics.
academic
this classification scheme is based on hemolytic reactions on BAP.
brown's
lancefield's classification scheme was pioneered by who?
Rebecca Lancefield
this classification scheme is based on cell surface carbohydrate Ag.
Lancefield's
how many serogroups are included in the lacefield classification scheme?
18
what are the sero groups in the lacefield classification scheme?
A to H
K to T
what are the clinically important sero groups in lacefield's classification scheme?
A
B
C
D
F
G
true or false
S. pneumoniae and many Viridans strep are not classified under lancefield.
true
this test if used in the ID of S. pyogenes and E. faecalis that produces L-pyrroglutamyl-aminopeptidase
PYR
PYR should be incubated at room temp for __ minutes before adding ___ drop of reagent.
2 minutes
1 drop
a red color within five minutes on a PYR indicates what result?
positive
orange or no red color on a PYR indicates what result?
negative
what are the QC organisms for PYR?
Positive: S. pyogenes or E. faecalis
Negative: S. agalactiae or E. mitis
the bacitracin (A or B disk) is used to identify which group of strep which is inhibited by a low concentration of bacitracin.
Grp A beta-strep
what amount of bacitracin is used in the A or B disk?
0.04 unit
in which disk test will any size zone of inhibition indicate suseptibility?
Bacitracin (A or B disk)
what are the QC organisms for Bacitracin (A or B disk).
Positive: S. pyogenes
Negative: S. agalactiae
the principle of the hippurate hydrolysis test is to ID organisms which produce the enzyme __________.
hippuricase
hippuricase can hydrolyze hippuric acid producing what two substances?
sdium benzoate
glycine
what is the substrate used in hippurate hydrolysis?
sodium hippurate
in what test do you inoculate 0.4 ml of 1% sodium hippurate reagent with 3-5 colonies of suspect organism. incubate for 2 hours at 35oC, then add 0.2 ml ninhydrin reagent and mix well?
hippurate hydrolysis
a deep purple color within 10 minutes on a hippurate hydrolysis test indicates what reaction?
positive
no color change on a hippurate hydrolysis test indicates what reaction?
negative
hippurate hydrolysis can be used as a standard test for what?
sodium benzoate
in order to test for sodium benzoate using hippurate hydrolysis, how long should the tue be incubated for before centrifugation?
20-24 hours
after centrifuging a hippurate hydrolysis to test for sodium benzoate, 0.2 ml of what reagent is used to interpret results?
ferric chloride
on the sodium benzoate hippurate hydrolysis test, what reaction does precipitate persiting indicate?
positve
if the solution is clear on a hippurate hydrolysis test for sodium benzoate, what result is indicated?
negative
what are the QC organisms for hippurate hydrolysis?
positive: S. agalactiae
negative: E. faecalis
what is the reference test for hippurate hydrolysis?
benzoate
hippurate hydrolysis takes greater than ___ hours to perform
20
the hemolysis accentuating factor is also called what?
CAMP factor
the hemolytic activity of staphylococcal beta-lysin on RBCs is enhanced by an extracellular factor produced by some beta-hemolytic streptococci is the principle of what test?
CAMP
in CAMP, perform a single steak of strep. isolate perpendicular to a beta-lysin producing strain of _________.
S. aureus
an arrow-head hemolysis at the junction of the two streaklines in CAMP indicates what reaction?
positive
abesence of arrow-head hemolysis on a CAMP test indicates what reaction?
negative
what are the QC organisms for CAMP?
Positive: S. agalactiae
Negative: E. faecalis
80% of what organism are CAMP (+)?
S. agalactiae
the principle of the optochin disk test is to presumptively identify what organism which is inhibited by a low concentration of optochin.
S. pneumoniae
what concentration of optochin is used in the optochin disk test?
5 mcg
ethylhydroxycupreine hydrochloride is also know as what?
optochin
what are the two optochin disks?
P disk
O disk
what are is the size of the optochin P disk?
6mm
what is the size of the optochin O disk?
10mm
optochin should be incubated at 35oC for 18-24 hours with what special reqiurement?
candle jar or CO2
what size zone of inhibition indicates susceptibility on a 6mm P disk?
>14 mm diameter
what size zone of inhibition indicates suseptibility on a 10 mm O disk?
>16mm diameter
what are the QC organisms for optochin?
positive: S. pneumoniae
negative: E. faecalis
the principle of the bile solubility test is to presumptively ID ___________ the cell of which are selectively lysed with a bile salt reagent.
S. pneumoniae
what is the reagent for the bile solubility test?
10% bile salts solution (sodium deoxycholate)
Bile solubilty should be incubated on a BAP at 35oC for how long?
30 min
if the colony disappears in the bile solubility test what is the reaction?
positive
if the colony remains intact on a bile solubility test what is the reaction?
negative
what are the QC organisms for bile solubility?
Positive: S. pneumoniae
Negative: S. viridans
Esculin hydrolysis is also known as what test?
BEM
to identify microorganism capable of growing in bile is the principle of what test?
Esculin hydrolysis (BEM)
the ability of an organism to hydrolyze esculin to glucose and esculetin with iron salt to for a ___________ complex, is the principle of esculin hydrolysis
brown-black
what does BEM stand for?
bile esculin medium
what is the color indicator in BEM?
ferric citrate
the blackening of more than half the slant in a esculin hydrolysis test indicates what reaction?
positive
no blackening or blackening of less than half the slant indiates what result in esculin hydrolysis?
negative
what are the QC organisms for esculin hydrolysis?
positive: group D enterococcus
negative: steptococcus or non-group D enterococcus
the principle of the test is to demonstrate bacterial grown tolerance to relatively high concentration of NaCl. It is used in differentiating group D enterococci and group D non-enterococci.
Salt tolerance test
what is the medium used in the salt tolerance test?
6.5% sodium chloride broth
turbidity w/without color indicates what reaction on a salt tolerance test?
positive
if the medium remains clear on a salt tolerance test, what is the reaction?
negative
what are the QC organisms used on the salt tolerance test?
Positive: group D enterococcus
Negative: group D non-enterococcus
the principle of this test is to detect S. pyogenes by Ag extraction. for use on direct specimen or pure culture.
rapid strep testing (grp. A)
what are the QC orgnaisms for rapid strep testing (grp. A)?
positive: S. pyogenes
negative: S. mitis
when diffrentiateing micrococcaceae organisms on a gram stain, staphylococci appear in grape like _______ while micrococci appear in ______.
staph: clusters
micrococci: tetrad
stapylococci and streptococcaceae grow rapidly in ___ hours while micrococci may take up to ___ hours to grow.