MacConkey Agars MacConkey Agar â¢ MacConkey ... - BVA Scientific

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MacConkey Agars, cont.User Quality ControlNOTE: Differences in the Identity Specifications and Cultural Response testing for media offered as both Difco and BBL brands may reflect differencesin the development and testing of media for industrial and clinical applications, per the referenced publications.MIdentity SpecificationsDifco MacConkey AgarDehydrated Appearance: Pink to pinkish beige, free-flowing,homogeneous.Solution:5.0% solution, soluble in purifiedwater upon boiling. Solution is reddishpurple, slightly opalescent.Prepared Appearance: Reddish purple, slightly opalescent.Reaction of 5.0%Solution at 25°C: pH 7.1 ± 0.2Difco MacConkey Agar BaseDehydrated Appearance: Pink to pinkish beige, free-flowing,homogeneous.Solution:4.0% solution, soluble in purifiedwater upon boiling. Solution is red,very slightly to slightly opalescent.Prepared Appearance: Red, slightly opalescent.Reaction of 4.0%Solution at 25°C: pH 7.1 ± 0.2Difco MacConkey Agar without Crystal VioletDehydrated Appearance: Pinkish beige, free-flowing, homogeneous.Solution:5.2% solution, soluble in purifiedwater upon boiling. Solution is reddishorange, clear to very slightlyopalescent.Prepared Appearance: Reddish orange, slightly opalescent.Reaction of 5.2%Solution at 25°C: pH 7.4 ± 0.2Difco MacConkey Agar without SaltDehydrated Appearance: Pinkish beige, free-flowing, homogeneous.Solution:4.7% solution, soluble in purifiedwater upon boiling. Solution is reddishorange, slightly opalescent.Prepared Appearance: Reddish orange, slightly opalescent.Reaction of 4.7%Solution at 25°C: pH 7.4 ± 0.2ContinuedIdentity SpecificationsBBL MacConkey AgarDehydrated Appearance:Fine, homogenous, may contain darkparticles.Solution:5.0% solution, soluble in purifiedwater upon boiling. Solution is mediumto dark, rose to brown-rose with orwithout a trace orange tint, clear toslightly hazy.Prepared Appearance: Medium to dark, rose to brown-rosewith or without a trace orange tint,clear to slightly hazy.Reaction of 5.0%Solution at 25°C: pH 7.1 ± 0.2BBL MacConkey Agar without Crystal VioletDehydrated Appearance: Fine, homogeneous, free of extraneousmaterial.Solution:5.2% solution, soluble in purifiedwater upon boiling. Solution is medium,red-orange to red-rose, slightly hazyto hazy.Prepared Appearance: Medium, red-orange to red-rose,slightly hazy to hazy.Reaction of 5.2%Solution at 25°C: pH 7.4 ± 0.2BBL MacConkey Agar without Crystal Violet or SaltDehydrated Appearance: Fine, homogeneous, free of extraneousmaterial.Solution:4.37% solution, soluble in purifiedwater upon boiling. Solution is medium,red-orange to red-rose, slightly hazyto hazy.Prepared Appearance: Medium, red-orange to red-rose,slightly hazy to hazy.Reaction of 4.37%Solution at 25°C: pH 7.4 ± 0.2ContinuedMacConkey Agar Base is prepared without added carbohydrates,which permits their addition either individually or in combination.It is recommended that carbohydrates such as sucrose orlactose be added in a concentration of 1% to the basalmedium.MacConkey Agar without Crystal Violet is a differentialmedium that is less selective than MacConkey Agar. The lackof crystal violet permits the growth of Staphylococcus andEnterococcus. Staphylococci produce pale pink to red coloniesand enterococci produce compact tiny red colonies either onor beneath the surface of the medium. The medium is usedalso to separate Mycobacterium fortuitum and M. chelonaefrom other rapidly growing mycobacteria. 9,10MacConkey Agar without Crystal Violet or Salt and MacConkeyAgar without Salt (which also lacks crystal violet) are differentialmedia used for isolating and cultivating gram-negativeenteric organisms and gram-positive cocci from waters, fecesand other sources suspected of containing these organisms, aswell as limiting the swarming of Proteus species.Principles of the ProcedurePeptones are sources of nitrogen and other nutrients. Lactoseis a fermentable carbohydrate. When lactose is fermented, alocal pH drop around the colony causes a color change in thepH indicator (neutral red) and bile precipitation. Bile salts,bile salts no. 3, oxgall and crystal violet are selective agentsthat inhibit growth of gram-positive organisms. Agar is thesolidifying agent.335
Section IIIM MacConkey Agars, cont.Cultural ResponseDifco MacConkey Agar or Difco MacConkeyAgar BasePrepare the medium per label directions. For MacConkey Agar Base,prepare without and with 1% added lactose. Inoculate and incubate at35 ± 2°C for 18-24 hours (and 40-48 hours for E. coli).INOCULUM COLONY BILEORGANISM ATCC CFU RECOVERY COLOR PPT.Enterococcus faecalis 29212 10 3 -2×10 3 Partial to – –completeinhibitionEscherichia coli 25922 10 2 -10 3 Good Pink to red; +w/o lactose:Colorless –Proteus mirabilis 12453 10 2 -10 3 Good Colorless –Salmonella choleraesuissubsp. choleraesuisserotype Typhimurium 14028 10 2 -10 3 Good Colorless –Difco MacConkey Agar without Crystal VioletPrepare the medium per label directions. Inoculate and incubate at 35 ±2°C for 18-48 hours.INOCULUM COLONY BILEORGANISM ATCC CFU RECOVERY COLOR PPT.Enterococcus faecalis 29212 10 2 -10 3 Good Red –Escherichia coli 25922 10 2 -10 3 Good Pink to red –Proteus mirabilis 12453 10 2 -10 3 Good Colorless –Salmonella choleraesuissubsp. choleraesuisserotype Typhimurium 14028 10 2 -10 3 Good Colorless –Difco MacConkey Agar without SaltPrepare the medium per label directions. Inoculate and incubate at 35 ±2°C for 18-48 hours.INOCULUM COLONY BILEORGANISM ATCC CFU RECOVERY COLOR PPT.Enterococcus faecalis 33186 10 2 -10 3 Good Red –Escherichia coli 25922 10 2 -10 3 Good Pink to red –Proteus mirabilis 12453 10 2 -10 3 Good Colorless, –no swarmingSalmonella choleraesuissubsp. choleraesuisserotype Typhimurium 14028 10 2 -10 3 Good Colorless –Shigella flexneri 12022 10 2 -10 3 Good Colorless –Cultural ResponseBBL MacConkey AgarPrepare the medium per label directions. Inoculate and incubate at35 ± 2°C for 18-24 hours (and 40-48 hours for E. coli).INOCULUM COLONY BILEORGANISM ATCC CFU RECOVERY COLOR PPT.Enterococcus faecalis 29212 10 3 -2×10 3 Partial to – –completeinhibitionEscherichia coli 25922 10 2 -10 3 Good Pink to +rose-redProteus mirabilis 12453 10 2 -10 3 Good Colorless –Salmonella choleraesuissubsp. choleraesuisserotype Typhimurium 14028 10 2 -10 3 Good Colorless –BBL MacConkey Agar without Crystal VioletPrepare the medium per label directions. Inoculate and incubate at35 ± 2°C for 18-24 hours and up to 48 hours if necessary (up to 11days for M. fortuitum).INOCULUM COLONY BILEORGANISM ATCC CFU RECOVERY COLOR PPT.Enterococcus faecalis 29212 10 3 -10 4 Good Rose-red –Escherichia coli 25922 10 3 -10 4 Good Pink to –rose-redMycobacteriumfortuitum 6841 10 3 -10 4 Good Rose-red –Salmonella choleraesuissubsp. choleraesuisserotype Typhimurium 14028 10 3 -10 4 Good Colorless –Staphylococcus aureus 25923 10 3 -10 4 Good Pink to –rose-redBBL MacConkey Agar without Crystal Violet or SaltPrepare the medium per label directions. Inoculate and incubate at35 ± 2°C for 18-24 hours and up to 48 hours if necessary.INOCULUM COLONY BILEORGANISM ATCC CFU RECOVERY COLOR PPT.Enterococcus faecalis 29212 10 3 -10 4 Good Rose-red –Escherichia coli 25922 10 3 -10 4 Good Pink to –rose-redProteus mirabilis 12453 10 3 -10 4 Good Colorless, –no swarmingSalmonella choleraesuissubsp. choleraesuisserotype Typhimurium 14028 10 3 -10 4 Good Colorless –336FormulaeDifco MacConkey AgarApproximate Formula* Per LiterPeptone ................................................................... 17.0 gProteose Peptone ....................................................... 3.0 gLactose ..................................................................... 10.0 gBile Salts No. 3 ........................................................... 1.5 gSodium Chloride ........................................................ 5.0 gAgar ......................................................................... 13.5 gNeutral Red ................................................................ 0.03 gCrystal Violet .............................................................. 1.0 mgDifco MacConkey Agar BaseConsists of the same ingredients without the lactose.BBL MacConkey AgarApproximate Formula* Per LiterPancreatic Digest of Gelatin ..................................... 17.0 gPancreatic Digest of Casein ........................................ 1.5 gPeptic Digest of Animal Tissue ................................... 1.5 gLactose ..................................................................... 10.0 gBile Salts ..................................................................... 1.5 gSodium Chloride ........................................................ 5.0 gAgar ......................................................................... 13.5 gNeutral Red ................................................................ 0.03 gCrystal Violet .............................................................. 1.0 mgDifco MacConkey Agar without Crystal VioletApproximate Formula* Per LiterPeptone ................................................................... 20.0 gLactose ..................................................................... 10.0 gBile Salts ..................................................................... 5.0 gSodium Chloride ........................................................ 5.0 gAgar ......................................................................... 12.0 gNeutral Red ................................................................ 0.05 g
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