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Kligler Iron Agar (KIA) and O/F Test Quiz
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Kligler Iron Agar (KIA) and O/F Test Quiz

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Questions and Answers

Which indicator is used in LIA to detect H2S production?

  • Phenol red
  • Bromocresol purple
  • Ferric ammonium citrate and sodium thiosulfate (correct)
  • Methyl red

    • What is the purpose of the slant and butt in LIA?

  • To detect hydrogen sulfide production
  • To detect glucose fermentation
  • To detect lysine decarboxylation
  • To detect aerobic and anaerobic metabolism (correct)

    • What is the significance of a purple slant and yellow butt on LIA?

  • Glucose fermentation positive, H2S production positive
  • Lysine decarboxylase negative, lysine deaminase positive
  • Lysine decarboxylase positive, lysine deaminase negative (correct)
  • Glucose fermentation negative, H2S production negative

    • Which test is used to detect the presence of $eta$-galactosidase activity?

    <p>Ortho-nitrophenyl-$eta$-D-galactopyranoside (ONPG) test</p> Signup and view all the answers

    Which agar medium is used to simultaneously detect glucose fermentation, lactose fermentation, and hydrogen sulfide production?

    <p>Triple Sugar Iron (TSI) Agar</p> Signup and view all the answers

    What color change is observed on the slant after adding FeCl3 if the result is positive for TRYPTOPHAN deamination?

    <p>Brown</p> Signup and view all the answers

    What is the color change observed in the UREASE test for rapid urease producers?

    <p>Bright pink</p> Signup and view all the answers

    Which of the following microorganisms is a slow/weak urease producer?

    <p>Enterobacter</p> Signup and view all the answers

    Which medium is commonly used to determine motility in microorganisms?

    <p>SIM (Sulfide Indole Motility) Medium</p> Signup and view all the answers

    What reaction indicates motility in the SIM (Sulfide Indole Motility) Medium?

    <p>Spread away from the stab line/hazy appearance</p> Signup and view all the answers

    Which of the following is NOT a member of ENTEROBACTERIACEAE that is motile?

    <p>Proteus</p> Signup and view all the answers

    What is the purpose of the Oxidation-Fermentation Tests (O/F tests)?

    <p>Identifying bacteria based on their oxidation and fermentation patterns.</p> Signup and view all the answers

    What happens in the Kligler iron agar tube for a non-oxidizer?

    <p>No acid is produced in both open and closed tubes.</p> Signup and view all the answers

    What is the purpose of Ortho-nitrophenyl-beta-D-galactopyranoside (ONPG)?

    <p>To determine the presence of slow lactose fermenters.</p> Signup and view all the answers

    Which color indicates an alkaline environment in the O/F medium?

    <p>Blue</p> Signup and view all the answers

    What does an oxidizer produce in the Kligler iron agar open tube?

    <p>Acid</p> Signup and view all the answers

    What is the pH indicator used in the Oxidation-Fermentation Tests?

    <p>Bromothymol blue</p> Signup and view all the answers

    Study Notes

    Lysine Iron Agar (LIA)

    • LIA is used in conjunction with TSI to screen stool specimens for enteric pathogens
    • Composition: aerobic slant, anaerobic butt, lysine, peptones, and a small amount of glucose
    • pH Indicator: BROMOCRESOL PURPLE
    • H2S Indicator: FERRIC AMMONIUM CITRATE & SODIARY THIOSULFATE
    • Manner of Inoculation: STAB-STREAK-STAB

    Reactions on LIA

    • Lysine Decarboxylation (butt/anaerobic): • K/K (Purple Slant/Purple Butt): LYSINE DECARBOXYLASE (+) LYSINE DEAMINASE (-) • K/A (Purple Slant/Yellow Butt): LYSINE DECARBOXYLASE (-) LYSINE DEAMINASE (-)
    • Note: Cadaverine neutralizes organic acids formed by glucose fermentation and reverts to alkaline state

    Oxidation-Fermentation Tests

    • Purpose: Identification of bacteria based on the oxidation-fermentation pattern
    • Medium: Hugh and Leifson O/F basal medium (OFBM)
    • pH Indicator: BROMOTHYMOL BLUE
    • Uninoculated: GREEN; Acid environment: YELLOW; Alkaline environment: BLUE
    • Two tubes: one overlaid with mineral oil (closed – anaerobic) and one without mineral oil (open – aerobic)
    • FERMENTER: ACID in BOTH open & closed tube
    • OXIDIZER: ACID in OPEN tube; no acid in closed tube
    • NON-OXIDIZER: NO ACID in BOTH open & sealed tube

    ORTHO-NITROPHENYL-BETA-D-GALACTOPYRANOSIDE (ONPG)

    • Determines the presence of Slow/Delayed/Late Lactose Fermenters (LLF)
    • ONPG is a colorless molecule structurally similar to LACTOSE
    • Can enter cell membrane without lactose permease and is converted into galactose and o-nitrophenyl (yellow chromogen) by Beta-galactosidase

    IMViC Reactions

    INDOLE TEST

    • Used to identify organisms that produce the enzyme TRYPTOPHANASE
    • Indicator: 10% FERRIC CHLORIDE (FeCl3) – added after incubation
    • Reactions: • (+) GREEN color on slant after adding FeCl3 • (-) Slant remains ORIGINAL COLOR after adding FeCl3

    UREASE TEST

    • Determines whether a microorganism can hydrolyze urea
    • Medium: Christensen’s Urea Agar or Stuart Urea Broth
    • pH Indicator: PHENOL RED
    • Reactions: • (+) BRIGHT PINK Coloration • (-) No Color Change (YELLOW)

    MOTILITY

    • Determines whether an organism is motile or non-motile
    • Medium: SIM (Sulfide Indole Motility) Medium or any semisolid motility medium
    • Streaking: STRAIGHT STAB
    • 1% triphenyltetrazolium chloride can be added to aid visualization
    • Reactions: • MOTILE (+): SPREAD AWAY from the stab line/HAZY APPEARANCE • NON-MOTILE (-): REMAIN at the stab line

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    Description

    Test your knowledge on Kligler Iron Agar (KIA) and the Oxidation-Fermentation (O/F) test. Learn about the purpose, medium composition, and pH indicator used in the O/F test. Identify bacteria based on their O/F pattern.

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