Identification flow charts

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., 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

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comma-shaped: Vibrio Campylobacter Spirillium Uniformly Bacillary: Coiled & sphero-plastic: Streptobacillus Fusobacterium Enterobacteriaceae Pseudomonas Aeromonas Alcaligenes Chromobacterium 2 .Identification flow charts Differentiation via Gram stains and cell morphology. Gram Stain & Morphological Flowchart Some Examples Gram Positive Cocci Bacilli Filamentous Round in clusters & tetrads: Staphylococcus Micrococcus Peptococcus Oval shape in Chains: Streptococcus Peptostreptococcus Enterococcus Club-shaped and/or in palisades: Corynebacterium Listeria Erysipelothrix Mycobacterium (Acid Fast) Propionobacterium Spore-bearing. uniform: Bacillus Clostridium Extensive branching: Actinomyces Arachnia Nocardia (partially Streptomyces acid-fast) Branching rudimentry or absent: Erysipelothrix Lactobacillus Eubacterium Gram Negative Cocci Bacilli Neisseria Veillonella Medium-sized cocco-bacillary: Acinetobacter Moraxella Tiny-coccobacillary: Brucella Bordetella Bacteroides Pleomorphic coccobacillary & filamentous: Exaggerated pointed ends: Haemophilus Bacteroides Pasteurella Francisella Actinobacillus Eikenella Cardiobacterium Flavobacterium Fusobacterium nucleatum Curved. large.

Lactobacillus spp. Mycobacterium spp. Lactobacillus spp. Acid Fast + Strict Anaerobes + You sure you have this? Go to Bergey's. - Clostridium spp.Identification flow charts Gram Positive Rods ID Flowchart Gram Positive Rods Bacillus spp. Activity + Corynebacterium kutsceri Corynebacterium xerosis Acid & Gas Acid Lactobacillus fermenti Lactobacillus casei Lactobacillus delbrueckii Mannitol + Lactobacillus casei Lactobacillus delbrueckii 3 . Clostrididium spp. Mycobacterium smegmatis Spore Forming + Bacillus spp. See separate flowchart. Mycobacterium spp. Clostridium spp Corynebacterium spp. Corynebacterium spp Lactobacillus spp. Bacillus spp. Starch Hydrolysis Glucose Ferm. Corynebacterium spp. Catalase + Corynebacterium spp. Lactobacillus spp.

marinus B. rest are neg. alvei B. licheniformis B. sphaericus + B. coagulans + B. pantothenticus Starch Hydrolysis (amylase) B. alcalophilus B. subtilis B. macquariensis B. stearothermophilus B. lentus B.) (acid via arabinose pos. thuringiensis B. marinus B. mycoides B. sphaericus Cell Diamenter ! 1"m (the width) (containing spore) Swollen Cell (containing spore) Swollen Cell + B. rest no) B. macquariensis (methyl red pos. cereus B. insolitus B.. rest are round) (Motile . licheniformis B. sphaericus B.) 4 .0) B. brevis B. brevis B. alcalophilus B. pasteurii B.) B. coagulans B. macquariensis B. polymyxa B. circulans B. licheniformis B. ID Flowchart Bacillus spp.) (catalase neg. megaterium (citrate pos. coagulans B. azotoformans B. thuringiensis B. macerans B. lentus B. subtilis (no growth at 55°C) (growth at 55°C) B. pasteurii B. firmus B. megaterium B. alvei B. popilliae VP + B. rest are) B.. badius B.) (manitol neg. popilliae B. cereus B.) (manitol pos. insolitus B. marinus (acid from glucose) (acid from glucose) B. circulans (acid via arabinose neg. laterosporus (round spore. macerans B. lentimorbus B. macerans B.) 6. cereus B. pantothenticus B.) + (add creatine as catalyst w/ reagents) B. megaterium B. insolitus B. brevis (methyl red neg. alvei + B. brevis B. larvae B.) (oval spore. badius B. macerans (gas from glucose pos.) B. subtilis B. subtilis B. lichenifomis B. alvei B.-rest pos. growth < pH 7. laterosporus B. badius (citrate neg. B. macquariensis B. pasteurii B. rest are oval) - Catalase Citrate B.5% NaCl Growth + B.) + B. circulans B. badius + + B. azotoformans B. no growth < pH 7. polymyxa B. mycoides B. thuringiensis B. pasteurii B. alcalophilus B. circulans B. macerans B. macquariensis B.0) (gas from glucose neg. polymyxa B. circulans B. alcalophilus B.) (gas from glucose neg. larvae B. alcalophilus B.It is this one ) Nitrate Reduction Citrate 6. pumilus (VP is pos. mycoides B.. subtilis B. larvae (gelatinase pos. popilliae (gelatinase neg. polymyxa B. coagulans B.Identification flow charts Bacillus spp. lentus B. pasteurii B.) B. pantothenticus B. coagulans Acid from Arabinose + B. schlegelii (grows at 55°C. pantothenticus B. sphaericus B. anthracis B. licheniformis B. brevis B.) (spore is not round.5% NaCl Growth + B. stearothermophilus B. anthracis B. circulans (growth at 55°C-rest neg. stearothermophilus B. azotoformans B. laterosporus B. anthracis (non-motile) (makes insecticidal protein) (colony has a rhizoidal appearance) B.

Micrococcus luteus Staphylococcus saprophyticus Optochin Sensitivity Yes Streptococcus pneumoniae No Streptococcus mitis 5 . Staphylococcus spp. (see Bergey’s) (growth with esculin hydrolysis) Catalase - Enterococcus faecalis Enterococcus faecium Enterococcus hirae + Enterococcus mundtii + Micrococcus spp.Identification flow charts Gram Positive Cocci ID Flowchart Gram Positive Cocci Micrococcus spp. Enterococcus spp. Streptococcus pneumoniae Streptococcus mitis Streptococcus pyogenes Group A Streptococcus spp. Growth w/ Tellurite Gamma (!) Mannitol Fermentation + Staphylococcus aureus Staphylococcus spp. Streptococcus spp. Hemolysis + Enterococcus faecalis Enterococcus faecium Enterococcus hirae (mannitol neg. Micrococcus spp.. Staphylococcus spp. yellow colony) Alpha (#) Staphylococcus spp. Bile Esculin Streptococcus spp.) (mannitol pos. (mannitol pos. Streptococcus pyogenes Group A Streptococcus spp. Group B Pyrrolidonearylamidase (PYR Test) Glucose Fermentation + Micrococcus varians Novobiocin Sensitivity Streptococcus pneumoniae Streptococcus mitis Yes Staphylococcus epidermidis Streptococcus pyogenes Group A + No Streptococcus spp. Streptococcus spp. Streptococcus spp. Group B (Streptococcus agalactiae) .. white colony) Enterococcus mundtii Beta (") Yellow Pigment (Colony) + Micrococcus spp. Enterococcus spp. Group B Enterococcus spp.

masoucida) (no gas from glucose) A. cichorii (To differentiate Biovars do a lecithinase test) (Lecithinase Pos. salmonicida A. orientalis V. syringae + P. syringae P. splendidius H2S Production Nitrate Reduction + A. salmonicida (subsp. cichorii P.Acid Pseudomonas spp. veronii 30°C Growth 37°C Growth + P. (Fluorescein)—use Psuedo F Agar + Vibrio spp. + Acid Glucose Fermentation . fischeri + V. orientalis V. veronii + V. hydrophila A. putida (Lecithinase Neg. logei V. chlororaphis (Colony Pigmented) P. fischeri V. putida P. fluorescens P. logei V. fischeri V. chlororaphis P. (See Bergey's) + Luminescent A. Vibrio Oxidase Test Enterobacteriaceae (See Separate Flowchartt) Vibrio spp.) 6 .. cichorii P. logei V. syringae (Oxidase Neg. (See Bergey's) Non-Fluorescent Diffusible Blue Pigment (Pyocyanin)—use Psuedo P Agar Motility Pigment P.) Other Pseudomonas spp. aeruginosa P. splendidius P. splendidius Other Vibrio spp. fluorescens P. (See Bergey's) P. aeruginosa (Yellow Colony) + A. + VP Other Aeromonas spp. orientalis V. masoucida) + V.) (Oxidase Neg.) + V. hydrophila A. Pseudomonas. Na+ Required for Growth Fluorescent Diffusible Yellow Pigment Aeromonas spp.Identification flow charts Gram Negative Rods ID Flowchart Gram Negative Rods + Aeromonas. hydrophila A. fluorescens P. veronii A. sobria (subsp. chlororaphis P. Aeromonas spp. putida P.

Serratia fonticola Klebsiella pneumoniae subsp. Red pigment Motile. No pigment + Citrobacter diversus (VP neg. pneumoniae Non-motile. none above are positive. ozaenae) Enterobacter aerogenes Enterobacter cloacae Enterobacter amnigenus Erwinia carotovora Serratia rubidaea Lysine Decarboxylase + Klebsiella pneumoniae subsp.Identification flow charts Family Enterobacteriaceae Lactose Positive ID Flowchart Family Enterobacteriaceae Lactose Fermentation + Citrobacter diversus Citrobacter freundii Enterobacter aerogenes Enterobacter cloacae Enterobacter amnigenus Enterobacter intermedius Erwinia carotovora Erwinia chrysanthemi Escherichia coli Klebsiella oxytoca Klebsiella pneumoniae Serratia fonticola Serratia rubidaea See Lactose Negative Flowchart for Enterobacteriaceae (subsp.) Escherichia coli Serratia rubidaea Enterobacter cloacae Acid from sorbitol Erwinia chrysanthemi Klebsiella oxytoca (VP and H2S pos. pneumoniae) +/Citrate Citrobacter freundii Serratia fonticola Klebsiella pneumoniae (subsp. No Pigment Motile.) Enterobacter amnigenus No acid from sorbitol H2S Production Enterobacter aerogenes ?? Erwinia carotovora Lysine unknown but Gelatinase positive. 7 .) (VP pos and H2S neg. ozaenae Motility neg. + Citrobacter freundii Motility pos. ozaenae and pneumoniae) Indole + Citrobacter diversus Escherichia coli Erwinia chrysanthemi Klebsiella oxytoca The Rest MR-VP +/+ Enterobacter intermedius -/+ Klebsiella pneumoniae (subsp.

neg. the rest pos. dysenteriae. flexneri) Indole Test Urease + Proteus mirabilis Proteus penneri Ornithine decarb. Serratia marcescens Red pigment Shigella sonnei Yersinia pestis Salmonella enterica KCN growth neg. flexneri) Shigella sonnei Yersinia enterocolitica Yersinia pestis Yersinia pseudotuberculosis Erwinia cacticida Proteus mirabilis Proteus penneri Salmonella bongori Salmonella enterica Serratia marcescens Serratia liquefaciens Shigella sonnei Yersinia pestis Yersinia pseudotuberculosis - Edwardsiella tarda Morganella morganii Proteus vulgaris Providencia stuartii + ?? Please See Bergey’s to ID: Shigella spp. (boydii. non-motile Erwinia cacticida Salmonella bongori Salmonella enterica Serratia marcescens Serratia liquefaciens Shigella sonnei Yersinia pestis Yersinia enterocolitica H2S Yersinia pseudotuberculosis + Edwardsiella tarda Proteus vulgaris Morganella morganii Providencia stuartii Erwinia cacticida Urease Ornithine Decarboxylase Motility + (lysine decarb. motile Ornithine decarb. Serratia liquefaciens No Red pigment (It is this one) 8 . pos.. neg. Ornithine decarb.. neg. (boydii.Identification flow charts Family Enterobacteriaceae Lactose Negative ID Flowchart Family Enterobacteriaceae Continued Lactose Negative Flow Chart Edwardsiella tarda Erwinia cacticida Morganella morganii Proteus mirabilis Proteus penneri Proteus vulgaris Providencia stuartii Salmonella bongori Salmonella enterica Serratia marcescens Serratia liquefaciens Shigella spp. dysenteriae..) Shigella sonnei Yersinia pestis + Proteus vulgaris Edwardsiella tarda + Providencia stuartii Morganella morganii Salmonella bongori Salmonella enterica Serratia marcescens Serratia liquefaciens Ornithine Decarboxylase H2S + + Salmonella bongori KCN growth pos.

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