FAMILY

ENTEROBACTERIACEAE
BY: PAUL AERON E. BANSIL, RMT
Enterobacteriaceae

◎Gram-Negative, Non-spore formers

◎Facultative anaerobic
◎Glucose Fermenters
◎Non Capsulated except Klebsiella and
Enterobacter
◎Reduces Nitrate to Nitrite
ALL MEMBERS OF THE FAMILY ENTEROBACTERIACEAE ARE MOTILE, EXCEPT

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 H2S POSITIVE ORGANISM
Salmonella, Proteus, Arizona, Citrobacter,
EDwarsiella

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ONLY OXIDASE POSITIVE
ENTEROBACTERIACEAE

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◎Isolated from Urinary Tracts (PEK)
- Proteus mirabilis
- Escherichia coli
- Klebsiella pneumoniae
• Associated with Diarrhea (SESY)
- Salmonella
- E. coli
- Shigella
- Yersinia 6
Escherichia

◎Escherichia coli
- Primary marker of fecal contamination in
water purification
- Leading of Nosocomial UTI
- TSI Reaction: A/A (+) Gas, H2S (-)
- IMViC: + + - -
- Dry Pink Colonies on MAC
- Green Metallic Sheen on EMB
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 E. coli Strains

◎Enteropathogenic E. coli (EPEC)

- Infantile Diarrhea
- Stool without Blood
◎Entertoxigenic E. coli (ETEC)
- Traveler’s Diarrhea
- Montezuma’s Revenge
- Watery Diarrhea w/o blood but with
Mucus
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 E. coli Strains

◎Enteroinvasive E.coli (EIEC)

- Shigella-like infection
- Watery Diarrhea with WBCs
◎Enterhemorrhagic E. coli (EHEC)
- 0157:H7
- Hemolytic Uremic Syndrome
- Bloody Diarrhea
◎Enteroadherent E.coli (EAEC)
- Watery Diarrhea 9
Klebsiella
◎Klebsiella pneumoniae
- Friedlandler’s Bacillus
- Causative agent of Community Acquired
Pneumonia
- Currant Jelly-Like Sputum
- Pink, Mucoid Colonies on MAC
- String Test (+)
- TSI A/A Gas (+), H2S (-)
- IMViC: - - + +
- KCN: + 10
Klebsiella

◎Klebsiella oxytoca
- Indole positive

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Enterobacter

◎Resembles Klebsiella
K. pneumoniae E. cloacae E. aerogenes

Lysine + - +
Decarboxylase

Arginine - + -
Decarboxylase

Ornithine - + +
Decarboxylase

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Serratia

◎Gelatinase, ONPG, Lipase, DNAse (+)

◎Prodigiosin
◎Serratia marcescens
- Bacteremic outbreaks in nurseries
- IMViC: - - + +
- TSI: K/A (+), H2S (-)

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Serratia

◎Serratia plymuthica
- Osteomyelitis following Motorcycle
Accidents
◎Serratia odorifera
- Potato-like Odor

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PAD TEST POSITIVE ORGANISMS?

PROTEUS
PROVIDENCIA
MORGANELLA

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Proteus

◎Rapid Urease Producer

◎Culture: Swarming Phenomenon
◎Burnt Chocolate Odor
◎Proteus vulgaris and Proteus mirabilis

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Providencia

◎Nosocomial outbreaks involving burned

patients
◎IMViC: + + - +
◎TSI: K/A Gas (-), H2S
◎Common Isolates: Providencia rettgeri,
Providencia stuartii

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Morganella

◎Morganella morganii
◎IMViC: + + - -
◎TSI: K/A Gas (+), H2S (-)

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Erwinia

◎Plant Pathogen

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Edwardsiella

◎Isolated from Cold blooded and warm

blooded animals
◎IMViC: + + - -
◎TSI: K/A Gas (+), H2S (+)

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Citrobacter

◎Resemble E.coli Colonies and

Biochemically resembles Salmonella
◎IMViC: - + - +
◎Grow on Simmon Citrate Agar

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Salmonella

◎Most serious pathogenic enterobacteria

for humans
◎Typhoid Fever
◎Acquired from improperly cooked
poultry, milk, eggs and dairy products

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Salmonella

◎Culture:
- Clear-Colorless Colonies (MAC)
- Colorless Colonies with Black Center
(SSA)
◎All are H2S positive except: S. paratyphi
◎All are Motile except: S. pullorum and S.
gallinarum

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Salmonella

◎ Specimens for SALMONELLA

Identification:
- Blood – 1st week of infection
- Urine – 2nd week of infection
- Stool – 3rd week of infection

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Shigella

◎Not a normal gastrointestinal flora

◎Non-Motile
◎Urease (-)
◎IMViC: - + - -
◎TSI: K/A
◎Shigella sonnei – stool without blood
◎Shigella dysenteriae – Bacillary Dysntery

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Yersinia

◎Yersinia pestis
- Class A Bioterrorism Agent
- Only Enterobacteriaceae with a vector
- Causative agent of the Bubonic Plague
- Stain: ________
- Microscopic Appearance: _______
- Culture: _______

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Yersinia

◎Yersinia enterocolitica
- Causative agent of waterborne
gastroenteritis
- Culture: _______ on CIN
- Selective Medium: Cefsulodin-Irgasan-
Novobiocin Agar
- Non-motile @ 37oC, Motile @ 22oC
- Requires cold enrichment technique
(4oC)
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Laboratory
Diagnosis
Culture Media

◎Mac Conkey
◎Mac Conkey with Sorbitol
◎XLD
◎HE
◎BSA
◎CIN

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 Mac Conkey
LACTOSE LATE-LACTOSE NON-LACTOSE
FERMENTERS FERMENTERS FERMENTERS
(EKE)
• E.coli • Citrobacter • Edwardsiella
• Klebsiella • Shigella sonnei • Proteus
• Enterobacter • Hafnia • Morganella
• Providencia
• Salmonella
• Shigella
• Yersinia
• Erwinia

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 Triple Sugar Iron

◎Determines whether a GN rod utilizes

Glucose, Sucrose or Lactose
◎1:10:10
◎pH indicator: ______
◎H2S indicators: ferrous sulfate and
sodium thiosulfate
◎18 to 24 Hours
◎Fermenter: Yellow
◎Non-Fermenter: Red
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 TSI Interpretation of Results

◎ Non-Fermenter (K/K)
- Alkaline slant/Alkaline butt
- Red Slant/Red Butt
- Not members of Family
Enterobacteriaceae
◎Sucrose Fermenter (K/A)
- Alkaline slant/Acid Butt
- Red Slant/Yellow Butt
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 TSI Interpretation of Results

◎Lactose and Sucrose Fermenter (A/A)

- Acid slant/Acid Butt
- Yellow Slant/Yellow Butt
◎Hydrogen Sulfide
- Formation of Black precipitate
◎Gas Production
- Formation of Bubbles, Cracks or Splitting
of Media
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 TSI Interpretation of Results
A/A G+ H2S - K/A G+ H2S- K/A G+ H2S+ K/A G- H2S-
• E.coli • Morganella • Proteus • Plesiomonas
• Klebsiella • Serratia • Salmonella • Providencia
• Enterobacter • Shigella
• C. koseri • C. freundii
• Edwardsiella • Yersinia

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 Lysine Iron Agar

◎Determine whether a GN decarboxylase

or deaminates lysine
◎Primarily important in differentiating
Salmonella and Citrobacter and PPM
◎Cadaverine and Putrescine
◎pH indicator: __________

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 Lysine Iron Agar

◎Deamination Occurs: BURGUNDY RED

◎Deamination Does NOT occur: PURPLE

◎Decarboxylation Occurs: PURPLE

◎Decarboxylation Does NOT occur:
YELLOW

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 Lysine Iron Agar
◎ Alkaline Slant/Alkaline Butt (K/K)
- Purple/Purple
- Lysine Deamination (–)/Lysine Decarboxylation (+)
◎ Alkaline Slant/Acid Butt
- Purple/Yellow
- Lysine Deamination (–)/Lysine Decarboxylation (-)
◎ Red Slant/Acid Butt
- Red/Yellow
- Lysine Deamination (+)/Lysine Decarboxylation (+)

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 IMViC

◎Indole
◎Methyl Red
◎Vogues-Proskauer
◎Citrate

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 IMViC

◎Indole Test
- Detects the production of indole from the
amino acid Tryptophan
- Reagent: Kovac’s Reagent
- Positive Result: Red or Pink Ring after
addition of Reagent

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 IMViC

◎Methyl Red
- Detects if the organism can produce acid
- Culture Media: MRVP broth/Peptone
Glucose Broth
- Positive Result: Red
◎Vogues-Proskauer
- Detects the formation of acetoin
- Reagent: 40% KOH and alpha-napthol
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 IMViC

◎Citrate Utilization Test

- Determines if the organism will utilize
citrate as sole source of carbon
- pH indicator: __________
- Culture Media: _________
- Positive Result: Green to Blue Slant

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 ONPG Test

◎ Orthonitrophenyl B-Galactopyranoside
- Detects late lactose fermenters
- Acting enzyme: ____________
- Positive Result: Yellow

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 Urease Test

◎Christensen’s Method
◎Rapid Urease Producers:____________
◎Reagent: Urea disk dissolve in 1mL
distilled water
◎Positive Result: Magenta or Pink

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