Bergey’s Manual PDF PDF

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Description

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

1

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, large, uniform: Bacillus Clostridium

Extensive branching: Actinomyces Arachnia Nocardia (partially acid-fast)

Branching rudimentry or absent: Erysipelothrix Lactobacillus Eubacterium

Streptomyces

Gram Negative Cocci

Neisseria Veillonella

Bacilli

Medium-sized cocco-bacillary: Acinetobacter Moraxella

Tiny-coccobacillary: Brucella Bordetella Bacteroides

Pleomorphic coccobacillary & filamentous: Haemophilus Bacteroides Pasteurella Francisella Actinobacillus Eikenella Cardiobacterium Flavobacterium

2

Exaggerated pointed ends: Fusobacterium nucleatum

Coiled & sphero-plastic: Streptobacillus Fusobacterium

Curved, comma-shaped: Vibrio Campylobacter Spirillium

Uniformly Bacillary: Enterobacteriaceae Pseudomonas Aeromonas Alcaligenes Chromobacterium

Identification flow charts

Gram Positive Rods ID Flowchart

Gram Positive Rods Bacillus spp. Clostrididium spp. Corynebacterium spp. Lactobacillus spp. Mycobacterium spp.

Mycobacterium smegmatis Spore Forming +

+

Corynebacterium spp. Lactobacillus spp. Mycobacterium spp.

Bacillus spp. Clostridium spp

Acid Fast

Strict Anaerobes

Corynebacterium spp Lactobacillus spp. -

+ Clostridium spp.

Bacillus spp.

You sure you have this? Go to Bergey's.

See separate flowchart.

Catalase

+

-

Corynebacterium spp.

Lactobacillus spp.

Starch Hydrolysis Glucose Ferm. Activity + Corynebacterium kutsceri

Acid

Acid & Gas

Corynebacterium xerosis

Lactobacillus fermenti

Lactobacillus casei Lactobacillus delbrueckii

Mannitol + Lactobacillus casei

3

Lactobacillus delbrueckii

Identification flow charts

Bacillus spp. ID Flowchart

Bacillus spp. B. subtilis B. cereus B. megaterium B. stearothermophilus B. polymyxa B. mycoides B. macerans B. thuringiensis B. macquariensis B. licheniformis B. lentus B. alvei B. anthracis B. alcalophilus B. coagulans B. brevis B. circulans B. firmus B. pantothenticus

Starch Hydrolysis (amylase)

+ VP

+ B. subtilis B. cereus B. polymyxa B. mycoides B. thuringiensis B. licheniformis B. alvei B. anthracis B. coagulans

-

(makes insecticidal protein)

B. mycoides

B. megaterium B. stearothermophilus B. macerans B. pantothenticus B. macquariensis B. lentus B. alcalophilus B. badius B. brevis B. circulans

(colony has a rhizoidal appearance)

B. popilliae

-

Catalase

(spore is not round, rest are)

Swollen Cell

Swollen Cell (containing spore)

-

B. stearothermophilus

B. megaterium

(growth at 55°C-rest neg.)

+

(citrate pos.)

B. macerans B. pantothenticus B. macquariensis B. lentus

B. badius (citrate neg.)

(oval spore, rest are round)

B. insolitus (acid from glucose)

B. marinus (acid from glucose)

Nitrate Reduction

6.5% NaCl Growth B. coagulans B. licheniformis B. subtilis

-

B. sphaericus B. pasteurii B. laterosporus

+

-

B. pasteurii

B. sphaericus

Citrate

+

B. larvae (gelatinase pos.) B. popilliae (gelatinase neg.)

B. azotoformans

(containing spore)

(catalase neg.-rest pos.)

(Motile - It is this one )

-

B. insolitus B. laterosporus B. pasteurii B. marinus B. sphaericus

B. alcalophilus B. brevis B. circulans

B. cereus

Citrate

+

+

B. badius

+

B. subtilis B. polymyxa B. lichenifomis B. alvei B. coagulans

(round spore, rest are oval)

B. popilliae B. pasteurii B. lentimorbus B. azotoformans B. badius B. insolitus B. larvae B. laterosporus

(add creatine as catalyst w/ reagents)

+ B. thuringiensis

B. azotoformans B. larvae B. pasteurii

(grows at 55°C, rest no)

(the width)

(non-motile)

(VP is pos., rest are neg.)

B. marinus B. sphaericus B. schlegelii

-

Cell Diamenter ! 1"m

B. anthracis

B. pumilus

B. polymyxa (manitol pos.)

B. alvei (manitol neg.)

-

+ B. macerans B. macquariensis B. alcalophilus B. brevis B. circulans

B. pantothenticus (acid via arabinose neg.)

B. circulans (acid via arabinose pos.)

6.5% NaCl Growth

+ B. subtilis

-

Acid from Arabinose

-

B. coagulans

(no growth at 55°C)

B. licheniformis

B. macquariensis

(growth at 55°C)

(methyl red pos.)

B. brevis (methyl red neg.)

4

B. macerans

+

(gas from glucose pos.)

B. alcalophilus (gas from glucose neg., no growth < pH 7.0)

B. circulans (gas from glucose neg., growth < pH 7.0)

Identification flow charts

Gram Positive Cocci ID Flowchart

Gram Positive Cocci

Catalase

-

+ Micrococcus spp. Staphylococcus spp.

Streptococcus spp. (see Bergey’s)

-

Micrococcus spp. Staphylococcus spp. Streptococcus spp. Enterococcus spp.

Bile Esculin (growth with esculin hydrolysis)

Streptococcus pneumoniae Streptococcus mitis Streptococcus pyogenes Group A Streptococcus spp. Group B Enterococcus spp. Streptococcus spp.

Enterococcus faecalis Enterococcus faecium Enterococcus hirae + Enterococcus mundtii

Enterococcus spp. Streptococcus spp.

Growth w/ Tellurite Gamma (!) Mannitol Fermentation + Staphylococcus aureus

+

Hemolysis

-

-

Enterococcus faecalis

Enterococcus faecium (mannitol pos., white colony)

Staphylococcus spp. 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)

Glucose Fermentation

Streptococcus pneumoniae Streptococcus mitis

Novobiocin Sensitivity

+ Micrococcus varians

+

Yes

Streptococcus pyogenes Group A

Staphylococcus epidermidis

- Micrococcus luteus

Streptococcus spp. Group B (Streptococcus agalactiae)

No Staphylococcus saprophyticus

Optochin Sensitivity

Yes Streptococcus pneumoniae

5

No Streptococcus mitis

Identification flow charts

Gram Negative Rods ID Flowchart

Gram Negative Rods Oxidase Test

+

Enterobacteriaceae (See Separate Flowchartt)

Aeromonas, Pseudomonas, Vibrio

+ Acid

Vibrio spp., Aeromonas spp.

-

- Acid

Pseudomonas spp.

Glucose Fermentation

Fluorescent Diffusible Yellow Pigment Na+ Required for Growth

(Fluorescein)—use Psuedo F Agar

Aeromonas spp.

-

+

+ VP

Vibrio spp.

-

+

Other Aeromonas spp. (See Bergey's)

A. hydrophila A. salmonicida

Luminescent

(subsp. masoucida)

P. aeruginosa P. chlororaphis P. cichorii P. fluorescens P. putida P. syringae

Other Pseudomonas spp. (See Bergey's)

(Oxidase Neg.)

A. sobria (no gas from glucose)

A. veronii

-

+ V. fischeri V. logei V. orientalis V. splendidius

Non-Fluorescent Diffusible Blue Pigment

Other Vibrio spp. (See Bergey's)

(Pyocyanin)—use Psuedo P Agar

Motility Pigment

+

(Yellow Colony)

A. hydrophila A. veronii

+

-

V. fischeri V. logei

V. orientalis V. splendidius

A. salmonicida (subsp. masoucida)

P. chlororaphis P. cichorii P. fluorescens P. putida P. syringae

+ P. aeruginosa

H2S Production Nitrate Reduction

+ A. hydrophila

30°C Growth

+

37°C Growth

+ V. fischeri

A. veronii

P. chlororaphis

+ V. logei

V. orientalis

(Colony Pigmented)

P. fluorescens

-

(To differentiate Biovars do a lecithinase test)

V. splendidius

6

P. putida (Lecithinase Neg.)

P. syringae (Oxidase Neg.)

P. cichorii (Lecithinase Pos.)

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

See Lactose Negative Flowchart for Enterobacteriaceae

(subsp. ozaenae and pneumoniae)

Serratia fonticola Serratia rubidaea

Indole The Rest + Citrobacter diversus Escherichia coli Erwinia chrysanthemi Klebsiella oxytoca

MR-VP

-/+

+/+

Klebsiella pneumoniae (subsp. pneumoniae)

Enterobacter aerogenes Enterobacter cloacae Enterobacter amnigenus Erwinia carotovora Serratia rubidaea

Enterobacter intermedius

+/Citrobacter freundii Serratia fonticola Klebsiella pneumoniae

Citrate

(subsp. ozaenae)

Lysine Decarboxylase -

+

+

-

Citrobacter diversus

Escherichia coli

Klebsiella pneumoniae subsp. pneumoniae Non-motile, No Pigment

(VP neg.)

Acid from sorbitol

Serratia rubidaea

Erwinia chrysanthemi

Enterobacter amnigenus

Motile, Red pigment

(VP and H2S pos.)

Enterobacter cloacae No acid from sorbitol

Enterobacter aerogenes

H2S Production

Klebsiella oxytoca (VP pos and H2S neg.)

Motile, No pigment

?? Erwinia carotovora Lysine unknown but Gelatinase positive; none above are positive.

-

+ Citrobacter freundii

Serratia fonticola Motility pos.

Klebsiella pneumoniae subsp. ozaenae Motility neg.

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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

Edwardsiella tarda Morganella morganii Proteus vulgaris Providencia stuartii

Serratia marcescens Serratia liquefaciens Shigella spp. (boydii, dysenteriae, 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

-

+

Indole Test Urease

+

?? Please See Bergey’s to ID: Shigella spp.

Erwinia cacticida Salmonella bongori Salmonella enterica Serratia marcescens Serratia liquefaciens Shigella sonnei Yersinia pestis

Proteus mirabilis Ornithine decarb. pos.

(boydii, dysenteriae, flexneri)

Proteus penneri

Yersinia enterocolitica

Ornithine decarb. neg., motile

H2S

Yersinia pseudotuberculosis Ornithine decarb. neg., non-motile

-

+ Edwardsiella tarda Proteus vulgaris

Morganella morganii Providencia stuartii

Motility

+

-

Erwinia cacticida

Shigella sonnei Yersinia pestis

(lysine decarb. neg., the rest pos.)

Urease

Salmonella bongori Salmonella enterica Serratia marcescens Serratia liquefaciens

Ornithine Decarboxylase

+

+

Proteus vulgaris

Providencia stuartii -

Edwardsiella tarda

Ornithine Decarboxylase

Morganella morganii

H2S

+ -

+ Salmonella bongori KCN growth pos.

Serratia marcescens Red pigment

Salmonella enterica KCN growth neg.

Serratia liquefaciens No Red pigment

(It is this one)

8

Shigella sonnei

Yersinia pestis...