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DILUTION OF BACTERIAL
SUSPENSION
BY: NELVEN M. GALLEGO, RMT, MLS(ASCPI)
MAAM GINA SADANG, RMT, MSMT
Exercise No. 13
Serial Dilution of
Bacterial Suspension
is a very significant
step in determining
bacterial colony count
NELVEN M. GALLEGO, RMT, MLS(ASCPi)
Exercise No. 13
Bacterial Suspension
Gram positive
microorganisms are normal
inhabitants of human skin &
mucous membranes
Infections caused by these
microorganisms can spread
through direct contact w/
infected person or fomites
NELVEN M. GALLEGO, RMT, MLS(ASCPi)
Exercise No. 16
BAP
Tryptones, soybean Differentiation of
digest, NaCl, agar, types of
5% blood:
Extracellular enzymes hemolysis
NELVEN M. GALLEGO, RMT, MLS(ASCPi)
Chocolate Agar Plate
CAP
5-10% Sheep, Warm enough to
lyse RBC and
rabbit, horse or release Hb and
human NAD
NELVEN M. GALLEGO, RMT, MLS(ASCPi)
Phenyl Ethyl Alcohol
PEA
G- rods may grow on PEA P. aeruginosa is not inhibited.
Some G+ cocci may require more
agar, but colonies are
than 24H of incubation. Although it
smaller than usual and contains blood it is not be used in
can be readily differentiated the interpretation of hemolytic
from those of G+ rods reactions
NELVEN M. GALLEGO, RMT, MLS(ASCPi)
Swarming of Proteus
Day1
CAP is incubated in an
anaerobic environment
using a candle jar.
Capnophiles
are microorganisms that thrive in the presence
of high concentrations of carbon dioxide
Obligate anaerobe
is any organism that does not
require oxygen for growth
NELVEN M. GALLEGO, RMT, MLS(ASCPi)
Anaerobic Cultivation
G+ cocci
Catalase
NELVEN M. GALLEGO, RMT, MLS(ASCPi)
Catalase test
Catalase
Positive Negative
Slide Tube
Coagulase Coagulase
NELVEN M. GALLEGO, RMT, MLS(ASCPi)
Procedure
Day2
Perform the tube
coagulase test by
inoculating on a tube
containing 0.5mL of human
plasma. Incubate at 37
degrees Celsius for 4 hours
Day2
Inoculate the
microorganism on
MSA. Incubate at 37
degree Celsius for
18-24 hours
NELVEN M. GALLEGO, RMT, MLS(ASCPi)
Mannitol Salt Agar
MSA
S. aureus can ferment This lowers the pH and
mannitol, the sole changes the color of
carbohydrate in the medium, the pH indicator,
to produce acid products phenol red, to yellow
NELVEN M. GALLEGO, RMT, MLS(ASCPi)
Procedure
MacConkey
LACTOSE NON-PATHOGENIC
FERMENTER ORANGE/ SALMON
ORANGE
NON- PATHOGENIC/ GREEN OR
LACTOSE BLUE COLOR
FERMENTER
NELVEN M. GALLEGO, RMT, MLS(ASCPi)
Thiosulfate Bile Salt Sucrose
TCBS
Bromthymol blue Sodium citrate, sodium
and, in some formulations, thiosulfate inhibits G
thymol blue are
incorporated to indicate +cocci and G-rods
the pH. normally present in stool
NELVEN M. GALLEGO, RMT, MLS(ASCPi)
Thiosulfate Bile Salt Sucrose
SSA
Bile salts, sodium
citrate, and brilliant Lactose is the sole
green, which inhibit the carbohydrate source in the
growth of G+ and many medium, and neutral red
LF, G- rods normally is the pH indicator
NELVEN M. GALLEGO, RMT, MLS(ASCPi)
Salmonella Shigella Agar
SSA
If an organism ferments
forming a black lactose, it will
precipitate in the produce acid and
center of the colony change the indicator to
pink-red
NELVEN M. GALLEGO,.RMT, MLS(ASCPi)
Salmonella Shigella Agar
XLD
A phenol red indicator decarboxylation of
in the medium detects lysine, which results in a pH
increased acidity from increase that causes the pH
carbohydrate. indicator to turn red
NELVEN M. GALLEGO, RMT, MLS(ASCPi)
Xylose Lysine Deoxycholate
BSA
the ferrous sulfate in which is deposited in the
this medium is reactive bacterial colony as a
with hydrogen sulfide black, insoluble
to produce ferric sufide precipitate
NELVEN M. GALLEGO, RMT, MLS(ASCPi)
Bismuth Sulfite Agar