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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
1
Identification flow charts
Gram Positive
Cocci Bacilli
Filamentous
Round in clusters Oval shape in Club-shaped and/or in Spore-bearing,
& tetrads: Chains: palisades: large, uniform:
Staphylococcus Streptococcus Corynebacterium Bacillus
Micrococcus Peptostreptococcus Listeria Clostridium Extensive Branching rudi-
Peptococcus Enterococcus Erysipelothrix branching: mentry or absent:
Mycobacterium (Acid Fast) Actinomyces Erysipelothrix
Propionobacterium Arachnia Lactobacillus
Nocardia (partially Eubacterium
acid-fast)
Streptomyces
Gram Negative
Cocci Bacilli
2
Identification flow charts
Gram Positive Rods ID Flowchart
Bacillus spp.
Clostrididium spp.
Corynebacterium spp.
Lactobacillus spp.
Mycobacterium spp.
Mycobacterium smegmatis
Spore Forming
+ - +
-
Strict Anaerobes Corynebacterium spp
Lactobacillus spp.
+ -
+ -
Corynebacterium spp. Lactobacillus spp.
Starch Hydrolysis
+ -
Mannitol
+ -
Lactobacillus casei Lactobacillus delbrueckii
3
Identification flow charts
Bacillus spp. ID Flowchart
Bacillus spp.
Starch Hydrolysis B. pumilus
(amylase) (VP is pos., rest are neg.)
B. subtilis
B. marinus
B. cereus - B. sphaericus
B. azotoformans
B. megaterium B. larvae
B. stearothermophilus B. schlegelii B. pasteurii
B. polymyxa
+ (grows at 55°C, rest no) (round spore, rest
B. popilliae are oval)
B. mycoides VP B. pasteurii B. popilliae
B. macerans (add creatine as catalyst B. lentimorbus
B. thuringiensis w/ reagents) -
B. azotoformans
B. macquariensis
B. badius
B. licheniformis
B. insolitus
B. lentus + - B. larvae
B. alvei
B. subtilis B. megaterium B. laterosporus Catalase Citrate
B. anthracis
B. alcalophilus B. cereus B. stearothermophilus
B. coagulans B. polymyxa B. macerans
B. mycoides B. pantothenticus + +
B. brevis
B. macquariensis B. badius -
B. circulans B. thuringiensis
B. licheniformis B. lentus (spore is not
B. firmus round, rest are) B. azotoformans B. larvae
B. pantothenticus B. alvei B. alcalophilus (gelatinase pos.)
B. badius B. insolitus
B. anthracis B. popilliae
B. brevis B. laterosporus
B. coagulans (gelatinase neg.)
B. circulans B. pasteurii
B. marinus
B. sphaericus
Cell Diamenter ! 1"m Swollen Cell Swollen Cell
(the width) (containing spore) (containing spore)
+ - + -
B. anthracis B. subtilis B. stearothermophilus B. megaterium + -
(non-motile) B. polymyxa (growth at 55°C-rest neg.) (citrate pos.)
B. thuringiensis B. lichenifomis B. macerans B. badius B. sphaericus B. insolitus
(makes insecticidal B. alvei B. pantothenticus (citrate neg.) B. pasteurii (acid from glucose)
protein)
B. coagulans B. macquariensis B. laterosporus B. marinus
B. mycoides B. lentus (oval spore, (acid from glucose)
(colony has a (catalase neg.-rest pos.) rest are round)
rhizoidal appearance) B. alcalophilus
B. cereus B. brevis
(Motile - It is this B. circulans
one ) Nitrate Reduction
Citrate + -
6.5% NaCl Growth B. pasteurii B. sphaericus
+ -
B. coagulans B. polymyxa + -
B. licheniformis (manitol pos.)
B. subtilis B. alvei B. macerans B. pantothenticus
(manitol neg.) B. macquariensis (acid via arabinose neg.)
B. alcalophilus B. circulans
B. brevis (acid via arabinose pos.)
B. circulans
B. macerans
+ - Acid from Arabinose
+ (gas from glucose pos.)
B. alcalophilus
B. subtilis B. coagulans - (gas from glucose neg.,
(no growth at 55°C) no growth < pH 7.0)
B. licheniformis B. macquariensis B. circulans
(growth at 55°C) (methyl red pos.) (gas from glucose neg.,
B. brevis growth < pH 7.0)
(methyl red neg.)
4
Identification flow charts
Gram Positive Cocci ID Flowchart
Mannitol Fermentation
+ -
- Hemolysis
+ Enterococcus faecalis Enterococcus faecium
Staphylococcus aureus Staphylococcus spp. (mannitol pos., white colony)
Micrococcus spp. Enterococcus hirae
(mannitol neg.)
Enterococcus mundtii
(mannitol pos., yellow colony)
Beta (")
Yellow Pigment (Colony)
Alpha (#)
Streptococcus pyogenes Group A
+ - Streptococcus spp. Group B
Micrococcus spp. Staphylococcus spp.
Pyrrolidonearylamidase
(PYR Test)
+
Micrococcus varians Yes +
Streptococcus pyogenes Group A
Staphylococcus epidermidis -
- - No Streptococcus spp. Group B
(Streptococcus agalactiae)
Micrococcus luteus Staphylococcus saprophyticus
Optochin Sensitivity
Yes No
5
Identification flow charts
Gram Negative Rods ID Flowchart
+ Acid - Acid
Vibrio spp., Aeromonas spp. Glucose Fermentation Pseudomonas spp.
+ -
+
VP P. aeruginosa Other Pseudomonas spp.
Vibrio spp. P. chlororaphis (See Bergey's)
- P. cichorii
+
P. fluorescens
A. hydrophila Other Aeromonas spp. P. putida
A. salmonicida (See Bergey's) P. syringae
Luminescent (subsp. masoucida) (Oxidase Neg.)
A. sobria
(no gas from glucose)
A. veronii
+ -
Non-Fluorescent Diffusible Blue Pigment
V. fischeri Other Vibrio spp. (Pyocyanin)—use Psuedo P Agar
V. logei (See Bergey's)
V. orientalis
V. splendidius Motility
- +
P. chlororaphis P. aeruginosa
- P. cichorii
Pigment + P. fluorescens
(Yellow Colony) A. hydrophila P. putida
A. salmonicida
A. veronii (subsp. masoucida) P. syringae
+ -
V. fischeri V. orientalis
V. logei V. splendidius
H2S Production
Nitrate Reduction
+ -
A. hydrophila A. veronii
+ -
30°C Growth 37°C Growth
P. putida
P. chlororaphis
+ + (Colony Pigmented)
(Lecithinase Neg.)
P. syringae
V. fischeri V. orientalis P. fluorescens (Oxidase Neg.)
- - (To differentiate Biovars
do a lecithinase test)
P. cichorii
V. logei V. splendidius (Lecithinase Pos.)
6
Identification flow charts
Family Enterobacteriaceae Lactose Positive ID Flowchart
Family Enterobacteriaceae
Lactose Fermentation
+ -
Indole
-
The Rest
+
Citrobacter diversus
Escherichia coli
Erwinia chrysanthemi
MR-VP
Klebsiella oxytoca +/+ -/+ Klebsiella pneumoniae
(subsp. pneumoniae)
Enterobacter intermedius Enterobacter aerogenes
Enterobacter cloacae
Enterobacter amnigenus
+/- Erwinia carotovora
Citrobacter freundii Serratia rubidaea
Citrate Serratia fonticola
Klebsiella pneumoniae
(subsp. ozaenae)
Lysine Decarboxylase
+ -
+ -
Klebsiella pneumoniae subsp. pneumoniae Enterobacter cloacae
Citrobacter diversus Escherichia coli Non-motile, No Pigment Acid from sorbitol
(VP neg.) Serratia rubidaea Enterobacter amnigenus
Erwinia chrysanthemi Motile, Red pigment No acid from sorbitol
(VP and H2S pos.) Enterobacter aerogenes
Klebsiella oxytoca H2S Production Motile, No pigment
(VP pos and H2S neg.)
??
Erwinia carotovora
Lysine unknown but Gelatinase
positive; none above are positive.
+ -
Citrobacter freundii Serratia fonticola
Motility pos.
Klebsiella pneumoniae subsp. ozaenae
Motility neg.
7
Identification flow charts
Family Enterobacteriaceae Lactose Negative ID Flowchart
+ Indole Test
Edwardsiella tarda
Morganella morganii
Urease
Proteus vulgaris
Providencia stuartii ?? + -
Please See Bergey’s to ID: Erwinia cacticida
Shigella spp. Proteus mirabilis
(boydii, dysenteriae, flexneri) Ornithine decarb. pos. Salmonella bongori
Yersinia enterocolitica Proteus penneri Salmonella enterica
Ornithine decarb. neg., motile Serratia marcescens
H2S Yersinia pseudotuberculosis Serratia liquefaciens
Ornithine decarb. neg., non-motile Shigella sonnei
Yersinia pestis
+ -
Edwardsiella tarda Morganella morganii
Motility
Proteus vulgaris Providencia stuartii
+ -
Erwinia cacticida Shigella sonnei
(lysine decarb. neg., the rest pos.) Yersinia pestis
Urease Ornithine Decarboxylase Salmonella bongori
Salmonella choleraesuis
Serratia marcescens
Serratia liquefaciens
+ +
Proteus vulgaris Providencia stuartii
Ornithine Decarboxylase
- -
Edwardsiella tarda Morganella morganii
H2S
+
Shigella sonnei -
+ -
Yersinia pestis
Salmonella bongori Serratia marcescens
KCN growth pos. Red pigment
Salmonella enterica Serratia liquefaciens
KCN growth neg. No Red pigment
(It is this one)