Bergey’s Manual of Determinative Bacteriology

1 Identification flow charts 1 Bergey s Manual of Determinative Bacteriology All of the unknowns will fall into the following groups in Bergey's Manual of Determinative Bacteriology (The pink book on the shelf in the laboratory). GROUP 4 Description: gram negative , aerobic /Microaerophilic rods and cocci Key differences are: pigments/fluorescent, motility, growth requirements, denitrification, morphology, and oxidase, read Genera descriptions Examples: Acinetobacter, Pseudomonas, Beijerinckia, Acetobacter GROUP 5 Description: Facultatively Anaerobic gram negative rods Key differences are: growth factors, morph., gram rxn., oxidase rxn., read Genera descriptions Examples: Family Enterobacteriaceae and Vibrionaceae GROUP 17 Description: gram -Positive Cocci Key differences are: oxygen requirements, morph.

2 , growth requirements (45 C and supplements), read Genera descriptions Examples: Micrococcus, Staphylococcus, Streptococcus, Enterococcus, Lactococcus, Aerococcus GROUP 18 Description: Endospore-Forming gram positive rods and cocci Key differences are: oxygen requirements, motility, morph, catalase Examples: Bacillus, Clostridium GROUP 19 Description: Regular, Nonsporlating gram positive rods Key differences are: morph., oxygen require, catalase Examples: Lactobacillus, Listeria GROUP 20 Description: Irregular, Nonsporlating gram -positive rods Key differences are: catalase, motility, morph., read Genera descriptions Examples: Actinomyces, Corynebacterium, Arthrobacter, Propionibacterium GROUP 21 Description: Weakly gram -Positive Nonsporlating Acid Fast Slender rods Key differences are: acid fast, growth Examples: Mycobacterium Identification flow charts 2 Differentiation via gram stains and cell morphology.

3 gram Stain & MorphologicalFlowchartSome ExamplesGram PositiveCocciBacilliRound in clusters & tetrads:StaphylococcusMicrococcusPeptoco ccusOval shape in Chains:StreptococcusPeptostreptococcusEn terococcusClub-shaped and/or in palisades:CorynebacteriumListeriaErysipe lothrixMycobacterium (Acid Fast)PropionobacteriumSpore-bearing, large, uniform:BacillusClostridiumBranching rudi-mentry or absent:ErysipelothrixLactobacillusEubact eriumTiny-cocco-bacillary:BrucellaBordet ellaBacteroidesNeisseriaVeillonellaExagg erated pointed ends:Fusobacterium nucleatumExtensive branching:ActinomycesArachniaNocardia (partially acid-fast)StreptomycesCurved, comma-shaped:VibrioCampylobacterSpirilli umPleomorphic coccobacillary & filamentous:HaemophilusBacteroidesPasteu rellaFrancisellaActinobacillusEikenellaC ardiobacteriumFlavobacteriumUniformly Bacillary:EnterobacteriaceaePseudomonasA eromonasAlcaligenesChromobacteriumFilame ntousGram NegativeCocciBacilliMedium-sized cocco-bacillary:AcinetobacterMoraxellaCo iled & sphero-plastic:StreptobacillusFusobacter iumIdentification flow charts 3 gram Positive rods ID Flowchart Bacillus sppMycobacterium smegmatisCorynebacterium sppLactobacillus sure you have this?

4 Go to Bergey' separate kutsceriLactobacillus delbrueckiiCorynebacterium xerosisLactobacillus caseiLactobacillus delbrueckiiLactobacillus caseiLactobacillus fermentiGram Positive RodsSpore FormingAcid FastStrict AnaerobesCatalaseStarch HydrolysisGlucose Ferm. ActivityAcid & GasAcidMannitol+------+++++Identificatio n flow charts 4 Bacillus spp. ID Flowchart B. subtilisB. cereusB. megateriumB. stearothermophilusB. polymyxaB. mycoidesB. maceransB. thuringiensisB. macquariensisB. licheniformisB. lentusB. alveiB. anthracisB. alcalophilusB. coagulansB. brevisB. circulansB. firmusB. pantothenticusB. subtilisB. cereusB. polymyxaB. mycoidesB. thuringiensisB. licheniformisB. alveiB. anthracisB. coagulansB. megateriumB. stearothermophilusB. maceransB. pantothenticusB.

5 MacquariensisB. lentusB. alcalophilusB. badiusB. brevisB. circulansB. badius (spore is not round, rest are)B. insolitusB. laterosporusB. pasteuriiB. marinusB. sphaericusB. pumilus (VP is pos., rest are neg.)B. marinusB. sphaericusB. schlegelii (grows at 55 C, rest no)B. popilliaeB. pasteuriiB. lentimorbusB. azotoformansB. badiusB. insolitusB. larvaeB. laterosporusB. azotoformansB. larvaeB. pasteurii(round spore, rest are oval)B. popilliaeB. subtilisB. polymyxaB. lichenifomisB. alveiB. coagulansB. stearothermophilus(growth at 55 C-rest neg.)B. maceransB. pantothenticusB. macquariensisB. lentus(catalase pos.)B. alcalophilusB. brevisB. circulansB. anthracis(non-motile)B. thuringiensis(makes insecticidal protein)B. mycoides(colony has a rhizoidal appearance)B.

6 Cereus(Motile - It is this one )B. megaterium(citrate pos.)B. badius (citrate neg.)B. pantothenticus (acid via arabinose neg.)B. circulans (acid via arabinose pos.)B. maceransB. macquariensisB. alcalophilusB. brevisB. circulansB. polymyxa (manitol pos.)B. alvei (manitol neg.)B. coagulansB. licheniformisB. subtilisB. subtilis (no growth at 55 C)B. licheniformis (growth at 55 C)B. coagulansB. larvae (gelatinase pos.)B. popilliae (gelatinase neg.)B. azotoformansB. sphaericusB. pasteuriiB. laterosporus(oval spore, rest are round)B. insolitus (acid from glucose)B. marinus (acid from glucose)B. pasteuriiB. sphaericusB. macerans (gas from glucose pos.)B. alcalophilus(gas from glucose neg., no growth < pH )B. circulans(gas from glucose neg.)

7 , growth < pH )B. macquariensis (methyl red pos.)B. brevis (methyl red neg.)Bacillus Hydrolysis(amylase)+VP(add creatine as catalyst w/ reagents)Cell Diamenter ! 1"m(the width)Swollen Cell(containing spore) NaCl GrowthAcid from ArabinoseNitrate ReductionCatalase-CitrateSwollen Cell(containing spore) NaCl Growth+++++++++++-----------Identificati on flow charts 5 gram Positive Cocci ID Flowchart Micrococcus aureusStaphylococcus variansMicrococcus luteusStaphylococcus saprophyticusStaphylococcus epidermidisStreptococcus pneumoniaeStreptococcus mitisStreptococcus pyogenes Group AStreptococcus spp. Group BEnterococcus faecalisEnterococcus faeciumEnterococcus hiraeEnterococcus mundtiiEnterococcus faecium(mannitol pos., white colony)Enterococcus hirae(mannitol neg.

8 Enterococcus mundtii(mannitol pos., yellow colony)Streptococcus spp.(see Bergey s)Enterococcus faecalisEnterococcus pyogenes Group AStreptococcus spp. Group BStreptococcus pneumoniaeStreptococcus mitisStreptococcus spp. Group B(Streptococcus agalactiae)Streptococcus pyogenes Group AStreptococcus pneumoniaeStreptococcus mitisGram Positive CocciCatalaseMannitol FermentationYellow Pigment (Colony)Glucose FermentationNovobiocin SensitivityBile Esculin(growth with esculin hydrolysis)Growth w/ TelluriteHemolysisOptochin SensitivityPyrrolidonearylamidase(PYR Test)YesNo+-++++++YesNo-------Gamma (!)Beta (")Alpha (#)Identification flow charts 6 gram negative rods ID Flowchart Aeromonas, Pseudomonas, VibrioEnterobacteriaceae(See Separate Flowchartt)Vibrio spp.

9 , Aeromonas fischeriV. logeiV. orientalisV. splendidiusA. hydrophilaA. salmonicida (subsp. masoucida)A. sobria (no gas from glucose)A. veroniiOther Vibrio spp.(See Bergey's)Other Aeromonas spp.(See Bergey's)V. fischeriV. logeiV. orientalisV. splendidiusA. hydrophilaA. veroniiA. salmonicida (subsp. masoucida)V. fischeriV. logeiV. orientalisV. splendidiusA. hydrophilaA. veroniiPseudomonas Pseudomonas spp.(See Bergey's)P. aeruginosaP. aeruginosaP. chlororaphisP. cichoriiP. fluorescensP. putidaP. syringae (Oxidase Neg.)P. chlororaphisP. cichoriiP. fluorescensP. putidaP. syringaeP. chlororaphis(Colony Pigmented)P. fluorescens(To differentiate Biovars do a lecithinase test)P. putida(Lecithinase Neg.)P. syringae(Oxidase Neg.)P. cichorii(Lecithinase Pos.)

10 gram negative RodsOxidase Test+-Glucose Fermentation+ Acid- AcidNa+ Required for GrowthLuminescentVPPigment(Yellow Colony)Motility30 C Growth37 C GrowthH2S ProductionNitrate ReductionNon-Fluorescent Diffusible Blue Pigment (Pyocyanin) use Psuedo P AgarFluorescent Diffusible Yellow Pigment(Fluorescein) use Psuedo F Agar--------+++++++++++---Identification flow charts 7 Family Enterobacteriaceae Lactose Positive ID Flowchart Citrobacter diversusCitrobacter freundiiEnterobacter aerogenesEnterobacter cloacaeEnterobacter amnigenusEnterobacter intermediusErwinia carotovoraErwinia chrysanthemiEscherichia coliKlebsiella oxytocaKlebsiella pneumoniae (subsp. ozaenae and pneumoniae)Serratia fonticolaSerratia rubidaeaSee Lactose negative Flowchart for EnterobacteriaceaeCitrobacter diversusEscherichia coliErwinia chrysanthemiKlebsiella oxytocaThe RestEnterobacter intermediusCitrobacter freundiiSerratia fonticolaKlebsiella pneumoniae (subsp.)