Scribd Logo
Upload

Welcome to Scribd!

  • 178 views

    Gram Negative Rods

    Uploaded by

    Anonymous pc5WTTgM
    This document provides information on the classification and identification of Enterobacteriaceae. It notes that Enterobacteriaceae are gram-negative rods that are oxidase-negative, facultative anaerobes capable of fermenting glucose with acid and gas production and reducing nitrates to nitrites. Key tests for identification include growth on MacConkey agar (lactose fermenters are pink, non-fermenters colorless), Salmonella-Shigella agar (lactose fermenters pink, Salmonella black centers from H2S), and triple sugar iron reactions (patterns indicate sugar fermentation and gas production). The document outlines the biochemical characteristics and reactions used to differentiate members of the Enterobacteriaceae

    Copyright:

    © All Rights Reserved
    Download as PPT, PDF, TXT or read online from Scribd

    Gram Negative Rods

    Uploaded by

    Anonymous pc5WTTgM
    178 views19 pages
    This document provides information on the classification and identification of Enterobacteriaceae. It notes that Enterobacteriaceae are gram-negative rods that are oxidase-negative, facultative anaerobes capable of fermenting glucose with acid and gas production and reducing nitrates to nitrites. Key tests for identification include growth on MacConkey agar (lactose fermenters are pink, non-fermenters colorless), Salmonella-Shigella agar (lactose fermenters pink, Salmonella black centers from H2S), and triple sugar iron reactions (patterns indicate sugar fermentation and gas production). The document outlines the biochemical characteristics and reactions used to differentiate members of the Enterobacteriaceae

    Original Description:

    A review and outline on gram negative rods (Microbiology)

    Copyright

    © © All Rights Reserved

    Available Formats

    PPT, PDF, TXT or read online from Scribd

    Did you find this document useful?

    Is this content inappropriate?

    This document provides information on the classification and identification of Enterobacteriaceae. It notes that Enterobacteriaceae are gram-negative rods that are oxidase-negative, facultative anaerobes capable of fermenting glucose with acid and gas production and reducing nitrates to nitrites. Key tests for identification include growth on MacConkey agar (lactose fermenters are pink, non-fermenters colorless), Salmonella-Shigella agar (lactose fermenters pink, Salmonella black centers from H2S), and triple sugar iron reactions (patterns indicate sugar fermentation and gas production). The document outlines the biochemical characteristics and reactions used to differentiate members of the Enterobacteriaceae

    Copyright:

    © All Rights Reserved
    Download as PPT, PDF, TXT or read online from Scribd
    Download as ppt, pdf, or txt
    178 views19 pages

    Gram Negative Rods

    Uploaded by

    Anonymous pc5WTTgM
    This document provides information on the classification and identification of Enterobacteriaceae. It notes that Enterobacteriaceae are gram-negative rods that are oxidase-negative, facultative anaerobes capable of fermenting glucose with acid and gas production and reducing nitrates to nitrites. Key tests for identification include growth on MacConkey agar (lactose fermenters are pink, non-fermenters colorless), Salmonella-Shigella agar (lactose fermenters pink, Salmonella black centers from H2S), and triple sugar iron reactions (patterns indicate sugar fermentation and gas production). The document outlines the biochemical characteristics and reactions used to differentiate members of the Enterobacteriaceae

    Copyright:

    © All Rights Reserved
    Download as PPT, PDF, TXT or read online from Scribd
    Download as ppt, pdf, or txt
    of 19

    Gram-negative rods

    Enterobacteriaceae

    Bacteria
    Gram positive
    Cocci

    Bacilli

    Gram negative
    Cocci

    Rods

    Characters of Enterobacteriaceae
    All Enterobacteriaciae

    Gram-negative rods
    Ferment glucose with acid production
    Reduce nitrates into nitrites
    Oxidase negative

    Facultative anaerobic
    Motile except Shigella and Klebsiella
    Non-capsulated except Klebsiella
    Non-fastidious
    Grow on bile containing media (MacConkey agar)

    Enterobacteriaceae
    Some Enterobacteriaceae are true pathogens

    Salmonella spp.
    Shigella spp.
    Yersinia spp.
    Certain strains of E. coli (ETEC, EPEC, EIEC, EHEC)

    Most members of the Enterobacteriaceae are


    opportunistic or cause secondary infections of
    wounds, the urinary and respiratory tracts, and
    the circulatory system e.g. E. coli.
    Enterobacteriaceae divided into TWO main
    groups according to action on LACTOSE
    Lactose Fermenters (LF)

    E. coli, Citrobacter, Klbesiella, Enterobacter

    Lactose Non-Fermenters (LNF)


    Salmonella, Shigella, Proteus, Yersinia

    Identification of Enterobacteriaceae
    Gram stain
    All Enterobacteriaceae are Gram-negative rods
    Arranged in single

    Identification of Enterobacteriaceae
    Biochemical reactions
    Oxidase test

    All members of Enterobacteriaceae are oxidase negative


    Pseudomonas is oxidase positive

    O/F test
    All members of Enterobacteriaceae are O+/F+
    Pseudomonas is O+/F-

    Nitrate reductase
    All members of Enterobacteriaceae are nitrate reductase positive
    Pseudomonas is nitrate reductase negative

    See & compare these tests under Pseudomonas Lab

    Classification of Enterobacteriaceae
    Enterobacteriaceae

    Lactose fermenters
    E. coli, Citrobacter,
    Klebsiella, Enterobacter

    Non-lactose fermenter
    Salmonell, Shigella
    Proteus, Yersinia

    There are several selective and differential media used to


    isolate distinguishes between LF & LNF
    The most important media are:
    MacConkey agar
    Eosin Methylene Blue (EMB) agar
    Salmonella Shigella (SS) agar
    In addition to Triple Sugar Iron (TSI) agar

    Differentiation between LF & NLF by Growth on MacConkey agar

    MacConkey agar is selective & differential medium for Enterobacteriaceae


    MacConkey Agar
    Contains

    Bile salts

    Crystal violet

    Inhibit growth of G+ve bacteria

    Cause of selectivity

    Lactose feremnters
    Pink colonies

    Lactose

    Cause of differential

    Neutral red

    pH indicator
    Acidic: Pink

    Lactose non feremnters


    colorless colonies

    Classification of Enterobacteriaceae according to


    lactose fermentation (growth on MacConkey Agar)
    Enterobacteriaceae

    Lactose Fermenters

    Neutral red

    Acid

    Pink colonies
    Escherichia coli
    Klebsiella spp
    Enterobacter spp
    Citrobacter spp

    Lactose Non-Fermenters
    No acid

    Colorless colonies

    Salmonella spp
    Schigella spp
    Proteus spp
    Yersinina spp

    Identification of Enterobacteriaceae
    Differentiation between LF & NLF by Growth on MacConkey agar
    Method:
    MacConkey agar is inoculated with tested organism
    using streak plate technique
    Incubate the plate in incubator at 37 C/24 hrs
    Results:
    LF organism appears as pink colonies (e.g. E. coli)
    NLF organism appears as colorless colonies (e.g.
    Shigella)
    Flame & Cool
    2

    3
    5

    Flame & Cool


    Flame & Cool

    Growth of Enterobacteriaceae on
    MacConkey agar

    Colorless colonies
    Uninoculated plate

    Lactose non feremters


    Salmonella, Shigella,
    Proteus

    Pink colonies
    Lactose feremters
    E. coli, Citrobacter
    Klebsiella, Enterobacter

    Reaction on Salmonella Shigella (SS) agar


    SS agar is a selective & differential medium used for isolation of
    Salmonella and Shigella
    The selective agents are bile salts, and brilliant green dye, which
    inhibit gram-positive organisms
    The medium contains only lactose as a differential agent and thus
    differentiates on the basis of lactose fermentation
    The formation of acid on fermentation of lactose causes the neutral red
    indicator to make pink colonies
    Non lactose fermenting organisms are colorless on the medium
    SS agar contains sodium thiosulfate and ferric ammonium citrate
    allows the differentiation of organisms that produce H2S
    Lactose fermenters, such as E. coli, have colonies which are pink
    Shigella appears transparent or amber
    Salmonella appears transparent or amber with black centers due to
    H2S production

    Lactose

    Lactose fermenter

    Acid

    H2S + Ferric ammonium citrate

    Neutral red

    Pink colonies
    Ferrous sulfide
    Black precipitate

    Identification of Enterobacteriaceae
    Differentiation between LF & NLF by Growth on SS agar

    Method:
    SS agar is inoculated with tested organism using
    streak plate technique
    Incubate the plate in incubator at 37 C/24 hrs
    1

    Flame & Cool


    2
    3
    4

    Flame & Cool

    5
    Flame & Cool

    Growth of Enterobacteriaceae on SS agar

    A. Klebsiella pneumoniae
    B. Escherichia coli
    C: Salmonella sp.
    D: Proteus mirabilis
    E: Ps. aeruginosa
    .

    Both are lactose fermenters


    Both Salmonella sp. & Proteus product H2S
    Pseudomonas colonies are nearly colorless

    Growth of Enterobacteriaceae on
    EMB agar

    Coli-type colonies are very dark,


    almost black e.g. E. coli

    Reaction on Triple Sugar Iron (TSI) Agar


    TSI contains

    Three different types of sugars


    Glucose (1 part)
    Lactose (10 part)
    Sucrose (10 part)

    Phenol red (acidic: Yellow)

    TSI dispensed in tubes with equal butt & slant


    Principle

    To determine the ability of an organism to attack a specific


    carbohydrate incorporated into a basal growth medium, with or
    without the production of gas, along with the determination of
    possible hydrogen sulphide production.

    Reaction on TSI
    Method:
    Inoculate TSI medium with an organism by
    inoculating needle by stabbing the butt and
    streaking the slant
    Incubate at 37C for 24 hours

    Reaction on TSI
    Butt color

    Slant color

    Yellow

    Red

    Yellow

    Red

    Yellow

    Yellow

    Red

    Red

    H2S

    Result
    s
    Result

    Negative
    Positive
    black in butt

    Negative

    Negative

    Example

    A/Alk/(Glucose fermented)

    LNF
    e.g. Shigella

    A/Alk/+
    (Glucose fermented with H2S)

    LNF
    e.g. Salmonella &
    Proteus

    A/A/(three sugars are fermented)

    LF
    e.g. E. coli, Klebsiella,
    Enterobacter

    Alk/Alk/(No action on sugars)

    Non fermenter e.g.


    Pseudomonas

    Practicle Work
    Gram stain
    Oxidase test
    O/F test
    Nitrate reductase test
    Growth on MacConkeys agar
    Growth on EMB agar
    Growth on SS agar
    Reaction on TSI

    You might also like