FAMILY ENTEROBACTERIACEAE

First proposed by Rahn in 1936

1972 - Edward & Ewing described the 11 Genera and 26

Species under the Family Enterobacteriaceae.

1985 - Farmer - noted 21 genera & 69 Species.

In the present we have more than 100 spp and over 30

genera.

BIOCHEMICAL DIFFERENTIATION
- Based on their TSI reaction

Family Enterobacteriaceae

Gram - Negative Bacilli Bacteria → Lactose Can be classified into Two (2) groups; Primary
Fermentation Pathogens and Opportunistic Pathogens.

Pink - Lactose Fermenter, bacteria color when ferments

using McConkey Agar and Eosine Methylene Blue.

Non-Lactose Fermenting Bacteria - Colorless

If does not ferment proceed to Oxidase Test

If Oxidase Positive = Pseudomonas

If Oxidase Negative → 𝑯𝟐 𝑺 Production Test

If there is production of 𝑯𝟐 𝑺 = color Black - Salmonella

and Proteus
Genera and Species to be considered
Gram(+) that produces 𝐻2 𝑆- Erisopelotrhix rhusiopathiea

CLASSIFICATION OF ENTERICS
Due to very large number of organisms in the Family
Enterobacteriaceae, species are grouped into TRIBES,
which have similar biochemical characteristics

Within each Tribe species are further sub-grouped under

genera and species.

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 VIRULENCE FACTOR
1. Pili - used for attachment; colonize the area and invade
tissues.

2. Some may possess PLASMID: Resistance to

antibiotics.
ESBLs (Extended Spectrum β-Lactamases) -
Bacteria that produces PLASMID
▪ E. coli, Klebsiella pneumoniae, Klebsiella
oxytoca
GENERAL CHARACTERISTICS
- Gram negative enteric coccobacilli, short, plump 3. Some may posses ANTIGENS that may be used to
bacilli identify other groups.
- Non - spore formers
- Facultative Anaerobes ANTIGENIC STRUCTURE
- Antigenic Group O or Somatic antigen
• Cell wall (O Ag or Somatic Ag) - somatic, - Found in the cell wall
HEAT STABLE, lipopolysaccharide - HEAT STABLE; LIPOPOLYSACCHARIDE
• Flagella (H Ag) - Flagellar, heat labile, protein - SEROTYPE 0111 - Diarrhea in INFANTS
• Capsule (K Ag) - capsular, heat labile, - SEROTYPE 0157 - Verotoxin (Verocytotoxin)
polysaccharide Production
o K1 - E. coli; Vi - S. typhi - For E. coli and Shigella serotyping

- BAP/CAP; Large moist gray colonies except K or Enveloped antigen

Klebsiella & Enterobacter. - Consists of capsular POLYSACCHARIDE
- HEAT LABILE
- All are γ- hemolytic except Escherichia coli - K antigen - E. coli
- Vi Antigen - S. typhi
- All are non-encapsulated except Klebsiella &
Enterobacter H or Flagellar antigen
Protein in nature
- All are GLUCOSE FERMENTERS and often with - Found in Flagellum or Flagella
gas production aerogenic except Shigella. - HEAT LABILE
- Found only among motile enteric
- All are MOTILE (Peritrichous Flagella; all over the - For Salmonella serotyping
bacilli) at 37°C except SKY.
• Shigella CULTURE MEDIAS
• Klebsiella
• Yersinia Pestis

❖ All yersinia are motile except Y. pestis at 25°C.

EMB
- All are Catalase Positive except S. dysenteriae

- All are Cytochrome Oxidase Negative except MAC

Plesiomonas c
XLD
CPON NO GF FA
HEA
- All are Nitrate Reducers except Erwinia, Pantoea
(Enterobacter) agglomerans, Photorabdus and
Xenorabdus. SSA

- Most are commensal flora of the intestinal tract except

BSA
Salmonella, Shigella and Yersinia.
- Some are with pili or fimbrae
- Grows well on McConkey Agar. TCBS
rd
Drug of choice: Aminoglycosides, Trimethoprim-SXT, 3
Gen Cephalosporins.

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EMB - Eosin Methylene Blue Agar
MAC - Mc Conkey Agar
XLD - Xylose Lysine Deoxycholate agar
HEA - Hectone Enteric Agar
SSA - Salmonella-Shigella Agar
BSA - Bismuth Sulfite Agar
TCBS - Thiosulfate Citrate Bile Salt Sucrose Agar
- Used for Vibrio spp.

MSA - used for Staphylococcus aureus

CTA - used for Neisseria
YerCINia Ashdown medium - used for Burkholderia
CIN - for Yesinia spp.
LIA: indicator; Bromcresol Purple
TSI: indicator; Phenol Red

Other Media:
a) GN Broth - For Salmonella
b) Selenite Broth - Enrichment broth for Salmonella and
Shigella
c) Tetrathionate Broth - Enrichment broth for
Salmonella
d) CIN (Cefsulodin-Irgasan Novobiocin) - for Yersinia
e) Brilliant Green Agar (BGA) - for other Salmonella
except S. typhi
- Inhibitory Agent - Brilliant Green; pH indicator -
Phenol Red

DIAGNOSTIC TEST FOR GRAM (-) ENTERIC BACILLI

a) TSI (Triple Sugar Iron)
- Uses TSI dispensed as slant and butt

Functions of TSI:
1. Detect ability of an organism to detect to ferment
CHO
2. Detects gas production
3. Detects 𝐻2 𝑆 Production

Composition:

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 Escherichia
Klebsiella
Enterobacter

b) LIA (Lysine Iron Agar)

Principle: Test to detect if the bacteria have the ability
to degrade amino acid lysine by DEAMINATION or
DECARBOXYLATION
- Can also detect 𝐻2 𝑆 production
Slant: Lactose & Sucrose
Description of reading:
Butt: Glucose
Slant (Aerobic) is observed for Deamination
Butt (Anaerobic) is observed for Decarboxylation
Fermentation of Glucose Process
Possible Result:
If cannot Ferment Glucose → (K/K)
pH indicator: Bromcresol Purple (BCP)
𝑯𝟐 𝑺 indicator: Ferric Ammonium Citrate = black
Glucose → Ferment → Pyruvic Acid (Glycolysis)
▪ Lysine decarboxylation - butt
If whole medium becomes Yellow (ACIDIC) →
• Positive - Purple; Negative - Yellow
Fermentation of Lactose /Sucrose → no change of color
(Yellow) (A/A) → Check if Produces 𝐻2 𝑆 → (A/A + 𝑯𝟐 𝑺)
▪ Lysine deamination - slant
• Positive - Red; Negative - Purple
If did not ferment Lactose/Sucrose → Deamination:
Peptone → 𝑵𝑯𝟐 → Red Slant (K/A) → if produces 𝑯𝟐 𝑺
Lysine DEAMINATION is an AEROBIC process
→ (K/A + 𝑯𝟐 𝑺)
which occurs on the SLANT of the media.
Deamination - Process of removing amino acid group in a
Lysine → Deamination → α-keto acid → reacts on
compound.
iron salt → red color

Deamination (+): Red (R)

Deamination (-): Purple (K)

Lysine DECARBOXYLASE is an ANAEROBIC

process which occurs in the BUTT of the media.

Lysine → Decarboxylase (Enzyme) →

CADAVERINE → Acid end product = Yellow

Cadaverine - Also has the ability to neutralize the acid

K/K end product that reverse it back to its alkaline state that
results to Color Purple (+)

K/A Decarboxylation (+): Purple (K)

Decarboxylation (-): Yellow (A)
A/A K/K

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Deaminase is an enzyme that removes the amino 𝑁𝐻2
group from the amino acids. 𝑁𝐻2 is alkaline

Decarboxylase is an enzyme that removes carboxyl

(COOH) group from the amino acids.

R/A

Proteus
Providencia
Morganella

YesCi

SLANT BUTT
DEAMINATION DECARBOXYLATION
R K K A
R P P Y
+ - + -

BACTERIAL METABOLISM
a) Mixed Acid Fermentation Test - Lactic acid,
succinic acid, formic acid, acetic acid; detected by
Methyl Red Test

b) Butanediol Fermentation Test - acetoin (acetyl

methyl carbinol) and butanediol which are neutral
end products; acetoin production is detected by
VOGES-PROSKAUER TEST

c) Butyric Acid Fermentation Test - butyric acid,

acetic acid, carbon dioxide and hydrogen
(Obligate anaerobes)

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IMViC Reaction of some CLINICALLY IMPORTANT e) UREASE Reaction
Bacteria

f) Malonate Utilization Test

(+) INDOLE (+) VP g) Gelatin Hydrolysis Test

May PEKPEC Po? KEESH
M - Morganella K - Klebsiella
P - Proteus vulgaris E - Enterobacter
E - Escherichia coli E - Ewingella
K- Klebsiella oxytoca S - Serratia
P - Providencia H - Hafnia
E - E. tarda
C - C. koseri
P - Plesiomonas

(+) Citrate
SaPro - ChEEK h) 𝑯𝟐 𝑺 Production
S - Salmonella enteriditis
P - Providencia
C - Citrobacter
H - Hafnia
E - Ewingella
E - Enterobacter
K - Klebsiella

Other Biochemical test

i) Motility Test

Yellow Β - Galactosidase

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j) Deaminase Reaction Test

k) Decarboxylation Reaction

l) KCN (Potassium Cyanide) Broth Test

m) MUG Test