mls micro competency #3

TSI set up

Pick up the center of an isolated colony, stab the butt of the slant and then streak the slant in a zigzag. Incubate with caps loose at 35 degrees celsius in a non-CO2 for 18-24 hours.

Urease set up

Pick up an isolated colony and inoculate onto the agar slant heavily over entire surface. Incubate at 35 degrees celsius in non-CO2 with caps loose loosenned for 18-24 hours.

Citrate set up

Pick up 1-2 colonies of organism and streak the slant then stab the butt. Inoculate with loosened cap aerobically (non co2) at 35 degrees Celsius for 24 hours

Motility test

Pick up an isolated colony and stab straight down medium halfway down, withdraw the needle along the same line of inoculation. Icubate tube at 35 degrees celsius in non-CO2 with loosened cap for 18-24 hours.

Aerococcus

Gram positive cocci, alpha hemolytic, catalase negative, PYR and NaCl positive, LAP negative. White dot in center of very small colonies.

Aerococcus

Strep pneumoniae

Gram positive cocci, alpha hemolytic, catalase negative, P disc (optochin) susceptible. Colonies are mucoid.

Strep pneumonia

TSI: alkaline slant (red)/no change

K/NC- no glucose, lactose or sucrose fermentation

TSI: acid slant (yellow)/acid butt (yellow)

A/A- glucose, lactose, and sucrose fermentation.

TSI: Acid slant (yellow)/acid butt (yellow) with gas

A/AG- Glucose, lactose and sucrose fermentation and gas producing

TSI: Alkaline slant (red)/acid butt (yellow)

K/A: Only glucose fermentation

TSI: Alkaline slant (red)/acid butt (yellow) With H2S

K/A with H2S- there is glucose fermentation and H2S production

LIA: Alkaline (purple)/Acidic (yellow)

K/A- lysine negative and glucose fermentation

LIA: Alkaline (purple)/ alkaline (purple)

K/K- lysine positive

LIA: Red/ acidic (yellow)

R/A- deamination of lysine and glucose fermentation

Urease positive

Hot pink slant or the whole tube

Urease negative

No change in the tube color (yellow)

K/A with gas and H2S

Salmonella TSI

K/K with gas and H2S

Salmonella LIA

Negative

Salmonella urease

K/A

Shigella TSI and LIA

Negative

Shigella urease

K/AG and H2S variable

Proteus TSI

K/A with H2S

Proteus LIA

Positive

Proteus urease

A/AG

Klebsiella pneumoniae TSI

Indole negative, MR negative (variable), urease positive, motility negative

Klebsiella pneumoniae

Indole positive, MR positive, urease negative, motility positive

E. Coli

A/AG

E. Coli TSI

Indole negative, MR positive, urease variable, motility positive

Citrobacter

A/AG with H2S

Citrobacter TSI

Indole positive, MR positive, urease positive, motility positive

Proteus vulgaris

K/AG and H2S variable

Proteus vulgaris TSI

Indole negative, MR positive, urease positive, motility positive

Proteus mirabilis

K/AG and H2S

Proteus mirabilis TSI

Colony morphology of proteus

MAC: flat, NLF BAP: thin film of swarming growth

Colony morphology of klebsiella

MAC: large, mucoid and lactose fermenter BAP: non-hemolytic, gray, mucoid

Colony morphology of E. Coli

MAC: flat, dry lactose fermenter BAP: beta or non-hemolytic gray colonies

Colony morphology of pseudomonas

MAC: flat, NLF, may be mucoid BAP: flat, irregular edge, gray green with silver sheen, beta hemolytic, can be mucoid. Odor: corn tortilla chips or over ripe grapes

Pseudomonas biochemical tests

Oxidase positive, catalase positive, TSI K/K silver sheen, motility top layer positive, growth on cetrimide agar- pigment is pyocyanin, growth at 42 degrees celsius, OF glucose positive

Acceptable urine culture

Shows significant colony counts of pathogenic organisms, and correlates with clinical symptoms

Contaminated urine culture

Shows mixed growth or evident nonpathogenic organisms

Quantitate using 10 microliter loop

Multiply the colony count by 100 cfu/ml

Klebsiella pneumonia

Proteus mirabilis

Klebsiella aerogenes

Serratia

Citrobacter freundii

Proteus vulgaris

Enterobacter cloacae

Pseudomonas

E. Coli

Shigella on HEK

Salmonella on MAC

Salmonella on HEK

Shigella on MAC

Shigella LIA

Proteus mirabilis LIA

Salmonella LIA