Variances Seen in Bacterial Analysis for Water - For Website 11-07

8/6/2019 Variances Seen in Bacterial Analysis for Water - For Website 11-07

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Variances seen inVariances seen inBacterial Analysis forBacterial Analysis for

Water and Waste WaterWater and Waste WaterSamplingSamplingGretchen HathawayGretchen Hathaway

Whatman Sales RepresentativeWhatman Sales RepresentativeJuly 19, 2007July 19, 2007


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TopicsTopics

MM--Endo (LTB, BGB, & ECB)Endo (LTB, BGB, & ECB)

MM--FC & MFC & M--FC with rosolic acidFC with rosolic acid

MI Broth and AgarMI Broth and Agar ColiCheck & ColiCheck with MUGColiCheck & ColiCheck with MUG


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MM--EndoEndo


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MM--EndoEndo

MM--Endo Broth is used for the enumeration of coliforms byEndo Broth is used for the enumeration of coliforms by

membrane filtration.membrane filtration.

Ideal for drinking water and waste water contaminantIdeal for drinking water and waste water contaminant

testing, Mtesting, M--Endo Coliform Broth is used to differentiateEndo Coliform Broth is used to differentiatelactose from nonlactose from non--lactose fermenting intestinal organisms.lactose fermenting intestinal organisms.


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MM--EndoEndo : Description: Description

MM--Endo is a red colored media, which needsEndo is a red colored media, which needsto be stored in the dark to preventto be stored in the dark to preventdiscoloration of the media.discoloration of the media.

GramGram--positive bacteria are inhibited on thispositive bacteria are inhibited on thismedia by the desoxycholate and lauryl sulfate.media by the desoxycholate and lauryl sulfate.The addition of ethanol increases theThe addition of ethanol increases theantibacterial nature of the formulation.antibacterial nature of the formulation.

Lactose fermenting organisms formLactose fermenting organisms formaldehydes, which react with Schiffs reagentaldehydes, which react with Schiffs reagent(basic fuchsin and sodium sulfite) to give red(basic fuchsin and sodium sulfite) to give redcolored zones around the colonies.colored zones around the colonies.

Coliform colonies are therefore red with aColiform colonies are therefore red with acharacteristic metallic sheen.characteristic metallic sheen.


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MM--Endo: InterpretationEndo: Interpretation

Production of both acid andProduction of both acid andaldehyde by lactose fermenters,aldehyde by lactose fermenters,

such assuch asEscherichia coliEscherichia coli, produce deep, produce deep

red colonies that color thered colonies that color the

surrounding medium and have asurrounding medium and have a

green metallic sheen.green metallic sheen.

NonNon--lactose fermenters formlactose fermenters form

colorless, translucent coloniescolorless, translucent colonies


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MM--Endo VerificationEndo Verification

The chart to the sideThe chart to the side

demonstrates the need ofdemonstrates the need of

verification steps due toverification steps due to

the type of growth.the type of growth.

Verification is done inVerification is done in

durham tubes with Lauryldurham tubes with Lauryl

Tryptose Broth, BrilliantTryptose Broth, Brilliant

Green Broth (to confirmGreen Broth (to confirmTotal coliform) and ECTotal coliform) and EC

Broth (to confirm FecalBroth (to confirm Fecal

coliform).coliform).


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ColiCheck andColiCheck andColiCheck with MUGColiCheck with MUG


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ColiCheck and ColiCheck with MUGColiCheck and ColiCheck with MUG

ColiCheckColiCheckis used for the presumptiveis used for the presumptive

identification of coliforms in water samples by aidentification of coliforms in water samples by a

presence/absence technique.presence/absence technique.

Two product types are available, ColiCheck thatTwo product types are available, ColiCheck that

identifies coliforms and ColiCheck with MUGidentifies coliforms and ColiCheck with MUG

that checks for the presence/absence ofthat checks for the presence/absence of

Escherichia coli.Escherichia coli.


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DescriptionDescription Bromocresol purple is a pH indicator which demonstrates aBromocresol purple is a pH indicator which demonstrates a

color change from purple to yellow in the presence of acid.color change from purple to yellow in the presence of acid.

Lactose fermenting organisms produce acid which initiatesLactose fermenting organisms produce acid which initiatesthe color change.the color change.

The presence of coliforms is detected with greaterThe presence of coliforms is detected with greatersensitivity by use of a relatively large sample volumesensitivity by use of a relatively large sample volume(100ml) in a single bottle.(100ml) in a single bottle.

The addition ofMUG (4The addition ofMUG (4--methylmethyl--umbellrifrylumbellrifryl----glucuronide) which is a fluorogenic enzyme allows theglucuronide) which is a fluorogenic enzyme allows themedia to selectively identifymedia to selectively identifyE. coliE. coli..

MUG is hydrolyzed by theMUG is hydrolyzed by theE. coliE. colispecific enzymespecific enzyme --glucuronidase to release 4glucuronidase to release 4--Methylumbelliferone whichMethylumbelliferone whichfluoresces under UV light (366nm).fluoresces under UV light (366nm).


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InterpretationInterpretation Yellow color in the sampleYellow color in the sampleafter 24 hours incubation atafter 24 hours incubation at3535 C indicates the presence ofC indicates the presence ofcoliform organisms.coliform organisms.

A red sample after 24 hourA red sample after 24 hourincubation at 35incubation at 35 C should beC should beincubated for further 24 hours.incubated for further 24 hours.

A red sample at 48 hoursA red sample at 48 hoursincubation at 35incubation at 35 C indicatesC indicatesno coliform organisms presentno coliform organisms present

in the sample.in the sample. ColiCheck w/ MUG:ColiCheck w/ MUG:

Florescence under UVLight isFlorescence under UVLight isspecific forspecific forE. coliE. coli..


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MM--FC brothFC broth


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MM--FCFC

MM--FC media (membrane Fecal ColiformFC media (membrane Fecal Coliform

media) is used for the detection of fecalmedia) is used for the detection of fecal

coliforms as an index of water pollution.coliforms as an index of water pollution.


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MM--FC: DescriptionFC: Description

Allows the developmentof fecal coliforms atelevated temperatures

(44.5 C).


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MM--FC: InterpretationFC: Interpretation

Bile salts included in themedium inhibit thegrowth of gram-positive

bacteria. Fecal coliforms ferment

lactose at elevatedtemperatures and

produce blue colonies. Other organisms form

grey to cream colonies.


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MM--FC with RosolicFC with RosolicAcidAcid


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MM--FC with Rosolic AcidFC with Rosolic Acid

M-FC Broth with Rosolic Acid is used in

waste water analysis. M-FC (Membrane Fecal

Coliform) media is designed for the detection of

fecal coliforms as an index of water pollution.


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MM--FC with Rosolic Acid: DescriptionFC with Rosolic Acid: Description

M-FC with Rosolic Acidacts and functions in thesame way as m-FC

Broth. Rosolic acid inhibits

bacterial growth ingeneral, except for fecal

coliforms.


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MM--FC with Rosolic Acid:FC with Rosolic Acid:

InterpretationInterpretation

Bile salts inhibit non-enteric bacteria.

Aniline blue indicates the

ability of fecal coliformsto ferment lactose to acidthat causes a pH changein the medium.

Other organisms formgrey to cream colonies.


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MI Broth and AgarMI Broth and Agar


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MI Broth and AgarMI Broth and Agar

Used for the simultaneous detection of total

coliforms andEscherichia coliin water

according to the Surface Water Treatment

Rule (USEPA) and the Total Coliform Rule

(USEPA), which applies to drinking water.


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MI Broth and Agar: DescriptionMI Broth and Agar: Description MI Broth detects the presence of coliformMI Broth detects the presence of coliform

bacteria by the production of bacteria by the production of --galactosidasegalactosidase,,which cleaves the substrate MUGal towhich cleaves the substrate MUGal toproduce 4produce 4-- Methylumbelliferone, whichMethylumbelliferone, whichfluoresces on exposure to UV light (366nm).fluoresces on exposure to UV light (366nm).

NonNon--coliforms do not produce this enzymecoliforms do not produce this enzymeand therefore do not fluoresce on theand therefore do not fluoresce on themedium.medium.

Escherichia coliEscherichia coliis detected by the compoundis detected by the compoundIBDG.IBDG.

The The --glucuronidaseglucuronidaseproduced byproduced byE. coliE. colicleaves the substrate to produce a bluecleaves the substrate to produce a blueindigo color in the colonies.indigo color in the colonies.

AsAsE. coliE. coliis also a total coliform, and alsois also a total coliform, and alsoproduces produces --galactosidase it will alsogalactosidase it will alsofluoresce.fluoresce.

The antibiotic cefsulodin is added to inhibitThe antibiotic cefsulodin is added to inhibitthe growth of gramthe growth of gram--positive bacteria andpositive bacteria andsome nonsome non--coliform gram negative bacteriacoliform gram negative bacteriathat can cause false positive reactions.that can cause false positive reactions.

Fewer False Positives are seenFewer False Positives are seenwith MI Broth.with MI Broth.


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MI Broth and Agar: InterpretationMI Broth and Agar: Interpretation

Fluorescent blue colonies areFluorescent blue colonies areEscherichia coliEscherichia coli..

Colonies that demonstrateColonies that demonstrateblue/white fluorescence where theblue/white fluorescence where thecolonies are clear, cream or palecolonies are clear, cream or paleyellow in color are other coliformyellow in color are other coliformorganisms.organisms.

Clear colonies that do not fluoresceClear colonies that do not fluoresceare nonare non--coliforms.coliforms.

Short incubation period PositiveShort incubation period Positivecontrol forcontrol forE. coliE. coliATCC 25922 andATCC 25922 andEnterobacter aerogenesEnterobacter aerogenesATCC 13048 can beATCC 13048 can beachieved in 18achieved in 18--24 hours at 3524 hours at 35C.C.

Negative control CompleteNegative control Completeinhibition ofinhibition ofPseudomonas aeruginosaPseudomonas aeruginosaATCC 10145 occurs at 35ATCC 10145 occurs at 35C after 24C after 24hours.hours.


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Other Products offered by WhatmanOther Products offered by Whatman

Monitors/ Analytical Filter FunnelsMonitors/ Analytical Filter Funnels

Membrane Systems and Membrane ButlerMembrane Systems and Membrane Butler

Dilution BottlesDilution Bottles

Petri dishes with padsPetri dishes with pads

Filter papersFilter papers Glass Fiber for Total Suspended and Volatile Solids, &Glass Fiber for Total Suspended and Volatile Solids, &TCLPTCLP

pH paperspH papers

Syringe filtersSyringe filters


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Variances Seen in Bacterial Analysis for Water - For Website 11-07

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