take an A4 page
divide into 3 parts
label each part as day 1, 2 and/or 3
provide subheadings, i.e macroscopy of specimen or culture, microscopy of specimen or culture, etc
in point form fill in everything that needs to be done
draw diagram if necessary
NB!!! you must be able to explain all types of inoculations/streaking/seeding
also all uses/purposes of media/biochemical tests used
expected results in biochemical tests
common morphology of orgs on all media used (include colours)
not sure if this helps. let me know.
Wednesday, February 9, 2011
urine processing DAY THREE
Read biochemical tests
the GNB was Klebsiella pneumoniae
citrate pos
indole neg (kovacs reagent)
H2S neg
non motile
acid slant and acid butt on TSI, gas pos
urease pos
PAD neg (10% FeCl2)
C albicans due to presence of germ tubes (no restriction between germ tube and yeast cell)
Enterococcus sp due to aesculin bile pos (black colour)and resistant to bacitracin
S saprophyticus due to coag neg, MSA pink colonies, DNA neg and Resistant to novobiocin
the GNB was Klebsiella pneumoniae
citrate pos
indole neg (kovacs reagent)
H2S neg
non motile
acid slant and acid butt on TSI, gas pos
urease pos
PAD neg (10% FeCl2)
C albicans due to presence of germ tubes (no restriction between germ tube and yeast cell)
Enterococcus sp due to aesculin bile pos (black colour)and resistant to bacitracin
S saprophyticus due to coag neg, MSA pink colonies, DNA neg and Resistant to novobiocin
urine processing DAY TWO Biochemical tests
Staphylococcus id
S saprophyticus is common cause of UTI in women and in men with indwelling catheters
only difference between S saprophyticus is its resistance to 5ug novobiocin
S saprophyticus is coag neg, MSA (pink colonies), DNA neg and novobioicin resistant.
so set up
DNA plate (stab and streak)
MSA (streak for single colonies)
coagulase (inoculate plasma with colony, incubate for up to 4 hours, remove and refridgerate if unable to read on same day)
seed CAB plate with suspension of org equivalent to McFarlands 0.5. Place novobiocin disc on plate)
GNB id
inoculate
TSI (stab and streak\
SIM
citrate *stab and streak)
PAD
urease
yeast id
set up germ tube
inoculate serum with colony
incubate at 37 degrees for up to 2 hours
make wet prep and examine for germ tubes which is pos for C albicans
Enterococcus id
inoculate BA plate. Place bacitracin disc in area of most growth (should be resistant)
inoculate aesculin bile (pos for Enterococcus)
S saprophyticus is common cause of UTI in women and in men with indwelling catheters
only difference between S saprophyticus is its resistance to 5ug novobiocin
S saprophyticus is coag neg, MSA (pink colonies), DNA neg and novobioicin resistant.
so set up
DNA plate (stab and streak)
MSA (streak for single colonies)
coagulase (inoculate plasma with colony, incubate for up to 4 hours, remove and refridgerate if unable to read on same day)
seed CAB plate with suspension of org equivalent to McFarlands 0.5. Place novobiocin disc on plate)
GNB id
inoculate
TSI (stab and streak\
SIM
citrate *stab and streak)
PAD
urease
yeast id
set up germ tube
inoculate serum with colony
incubate at 37 degrees for up to 2 hours
make wet prep and examine for germ tubes which is pos for C albicans
Enterococcus id
inoculate BA plate. Place bacitracin disc in area of most growth (should be resistant)
inoculate aesculin bile (pos for Enterococcus)
urine processing DAY TWO
examine antimicrobial plate
any zone around disc indicates that the px is on antibiotics and will influence the significance of any growth of organisms
examine CLED plate
estimate the amount of growth
growth up to the squiggle in the middle of plate equals 100 000 orgs per ml of urine
growth up to a level below the squiggle decreases by a power of 10. (please see me if you dont understand this)
examine the BA Growth should represent totality of all orgs seen on other plates
examine MaC for LF/NLF (LF are pink, NLF are brown, transparent, colourless)
examine SAB for yeasts (distinctive odour)
NB! you must get used to the smell of bacteria. Orgs like yeasts, E coli, Klebs, S aureus have very distinct smells. Smells is one way of guiding you to a possible identity of the organism.
set up biochemical tests to identify any potential pathogens seen.
contd in next post
any zone around disc indicates that the px is on antibiotics and will influence the significance of any growth of organisms
examine CLED plate
estimate the amount of growth
growth up to the squiggle in the middle of plate equals 100 000 orgs per ml of urine
growth up to a level below the squiggle decreases by a power of 10. (please see me if you dont understand this)
examine the BA Growth should represent totality of all orgs seen on other plates
examine MaC for LF/NLF (LF are pink, NLF are brown, transparent, colourless)
examine SAB for yeasts (distinctive odour)
NB! you must get used to the smell of bacteria. Orgs like yeasts, E coli, Klebs, S aureus have very distinct smells. Smells is one way of guiding you to a possible identity of the organism.
set up biochemical tests to identify any potential pathogens seen.
contd in next post
urine processing DAY ONE
day one
macroscopy of specimen
describe odour (if applicable) colour, turbidity, volume
microscopy of specimen
decant few ml into a centrifuge tube
centrifuge and make wet prep with sediment
examine using up to 40X obkective
examine for cells, crystals, casts, and bacteria/yeasts
if casts and crystal seen, name type (significant as it denotes type of kidney damage)
culture of specimen
use plates dependent on what seen in wet prep (use urine from original urine specimen container)
include BA (enriched medium, usually all orgs grow), Mac for LF/NLF (GNB), CLED (total colony count, streak for single colonies without flaming, use 1/1000 ul loop)
include SAB if yeasts seen in wet prep
ALWAYS remember potential pathogens found in urine
perform an antimicrobial test to dtermine if px is on antimicrobial therapy
make a suspension of B subtilis equivalent to McFarlands 0.5
seed a CAB plate using the suspension
place a sterile filter paper disc on plate
place a loopful (1/100 ul) of urine on disc
incubate all plates at 37 degrees, aerobically for 18-24 hours
lastly do a dipstick to determine any abnormal chemistry.
NB! Nitrites indicate bacterial activity i.e. reduction of nitrates
macroscopy of specimen
describe odour (if applicable) colour, turbidity, volume
microscopy of specimen
decant few ml into a centrifuge tube
centrifuge and make wet prep with sediment
examine using up to 40X obkective
examine for cells, crystals, casts, and bacteria/yeasts
if casts and crystal seen, name type (significant as it denotes type of kidney damage)
culture of specimen
use plates dependent on what seen in wet prep (use urine from original urine specimen container)
include BA (enriched medium, usually all orgs grow), Mac for LF/NLF (GNB), CLED (total colony count, streak for single colonies without flaming, use 1/1000 ul loop)
include SAB if yeasts seen in wet prep
ALWAYS remember potential pathogens found in urine
perform an antimicrobial test to dtermine if px is on antimicrobial therapy
make a suspension of B subtilis equivalent to McFarlands 0.5
seed a CAB plate using the suspension
place a sterile filter paper disc on plate
place a loopful (1/100 ul) of urine on disc
incubate all plates at 37 degrees, aerobically for 18-24 hours
lastly do a dipstick to determine any abnormal chemistry.
NB! Nitrites indicate bacterial activity i.e. reduction of nitrates
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