ENTEROBACTERIACEAE

Dr. Esquivel

(4) Klebsiellae
Genus: Klebsiella
Species: K. pneumonia nosocomial pneumonia
K. azaenae
K. rhinosclenmatis
K. oxytoca

A. Bergys Classification
1. Escherichiacheae
2. Klebsiellae
3. Proteae
4. Yesiniae
5. Erwinieae
B. Edwards and Ewings Classification (Most accepted)
1. Escherichiacheae
2. Edwardsiellae
3. Salmonellae
4. Klebsiellae
5. Proteae
6. Yersinieae
7. Erwineae

Genus: Enterobacter
Species: E. cloacae
E. aerogenes
E. aglomerans
E. sakazakii
E. gergoviae
Genus: Serratia
Species: S. marcescens
S. liquifaciens
S. rubideae red pigment

Edwards and Erwings Classification

(1) Escherichiacheae
Genus: Escherichia
Species: E. coli
E. hermanii
E. vulneris

Genus: Hafnia
Species: H. alvei
(5) Proteae
Genus: Proteus
Species: P. vulgaris UTI-related cather-related
P. mirabilis
P. rettgeni
P. penreri

Genus: Shigella
Species: S. dysenteriae bacterial dysentery
S. flexneri
S. boydii
S. sonnei

Genus: Providencia
Species: P. alcalifaciens
P. stuartii

(2) Edwardsiella
Genus: Edwardsiellae
Species: E. tarda
(3) Salmonellae
Genus: Salmonella
Species: S. typhi typhoid fever
S. cholera-suis
S. enteritidis
Genus: Arizone
Species: A. hinshawii

Genus: Morganella
Species: M. morganni
(6) Yersinieae
Genus: Yersinia
Species: Y. pestis plague
Y. pseudotuberculosis
(7) Erwineae
Genus: Erwinia
Genus: Rectobacterium
Genus: Tatumella
Species: T. ptyseus

Genus: Citrobacter
Species: C. freundii
C. diversus

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1. Microscopic
- urine, blood, stool, CSF, pus
CHARACTERISTICS
- gram (-) rods
- non spore forming
- facultative ANaerobe
- grow in simple media
- ferment glucose and produce acid
- motile: some have peritrichous flagella (Pseudomonas and
Vibrio), others have polar flagella; non-motile: Shigella,
Klebsiella)
- some have capsule (K. pneumonia)
- some have slime layer (E. coli)
- most are piliated
- reduce nitrate to nitrite
- H2S production is variable (S. typhi)
- Some have extrachromosomal elements that code for
production of bacteriocins or colicins that kill other
organisms
- destroyed easily by heat and disinfectants
- tolerates cold tem and sensitive to drying.
DETERMINANTS OF PATHOGENICITY

2. Culture and isolation

- colonial morphology
Typical colonies:
a. large gray, watery, mucoid colonies
- capsulated organisms (K. pneumoniae)
b. thin film/waves with swarming growth
- motile organisms (Proteus)
c. pigmented colonies
- green metallic sheen in EMB (E. coli)
- pink colonies on SSA by E. coli
- non-pigmented nonlactose fermenters pathogenic
(Salmonella and Shigella)
3 general types of culture media
a. nonselective media for isolation
-BAP
b. selective and differential culture media.

Major antigens
* selective only gram negative
- XLD (Xylose Lysine Desoxycholate)
- HA (Hektoen Agar)

1. K antigen polysaccharide capsule

2. H antigen flagellar antigen
3. O antigen somatic antigen

* selective and differential

- SSA
- EMB
- MacConkey Agar

Virulence Factors:
1. endotoxin LPS
- Polysaccharide core
- destroys cell
- O antigen (immunogenic)
- Lipid A fever production
2. exotoxins
- Shiga toxin (S. dysentery)
- Choleragen (V. cholerae)
Adhesion Colonization Factors
1. Pili
2. O antigen
3. Other membrane proteins
:Capsule and other protective surface antigen

c. Enrichment Broth
- Selenite F in HA: 8-12 hours
- Gram Negative broth in HA: 4 hours
* incubate all plates @ 35-37 degrees for 24-48 hours
3. Biochemical Testing:
a. H2S production
black pigment H2S prodn in TSI (butt/slant)
Salmonella
b. utilization of carbohydrate
- lactose, glucose, sucrose
- TSIA or KIA
- TSIA 1: red yellow = acid production

LABORATORY DIAGNOSIS

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- TSIA 2: crack gas production

- attachment and penetration of intestinal epithelium

- target: Ileum (peyers patches)

c. IMViC Test
c.1. INDOLE INDOLE prodn - SIM medium
c.2. METHYL RED RXN ACID prodn RED!!
c.3. VOGES-PROSKAUER ACETOIN
production RED!
c.4. CITRATE UTIL. CITRATE util. SCA GREEN

- ANTI-PHAGOCYTIC FACTORS:
-a. catalases and superoxide dismutase:
Both can neutralize active oxygen
-b. definsin:
- small cationic proteins that facilitate bacterial
killing by phagolysosome.

d. Urease Production
- Urea Agar Slant
- Urea Ammonia + CO2

-Vi or VIRULENCE ANTIGEN

- anti phagocytic activity

e. Decarboxylation of Lysine, Ornithine and Arginin

ANTIGENIC STRUCTURES

f. Production of Phenylalanin

1. H antigen
- inactivated by heating of 60 degress,alcohols and acids
-best prepared for serological testing by addition of
formalin to gram negative motility broth.

4. Motility study
1. Agar block technique
2. Hanging drop
3. Semi-solid medium
SALMONELLA
- nonlactose fermenters
- colorless colonies
- gas prodn when fermenting glucose
- H2S produces from thiosulfate
- motile: peritrichous
Species:
1. S. typhi / Eberths Bacillus
2. S. paratyphi A.
3. S. paratyphi B or schottmuelleri
4. S. paratyphi C or hirsshfeldi
5. S. cholera-suis
6. S. tymimurum
7. S. enteritidis / Gartners Bacillus
S. typhi
- gram 9 (-)
- NON encapsulated
- NON-spore forming
-motile: peritrichous
- resistant to:
* Brilliant Green
* Sodium tetrathionate
* Sodium desoxycholate
DETERMINANTS OF PATHOGENICITY
- Endotoxin

2. O antigen
- surface
- resistant at 100 degrees,alcohol and diluted acids
- non-motile with treatment, heat and alcohol
- LPS-free proteins
3.Vi antigen
- destroyed by heat for 1 hour at 60 degrees, acids &
phenol
- present in virulent strains
DISEASE: SALMONELLOSIS
1. Enterocolitis
- S. enteritidis, S. typhimurium
- self-limited,lasting 2-5 days
-diarrhea
- Cx: dehydration and electrolyte imbalance
2. Osteomyelitis
- S. enteritidis with sickle cell anemia
- bone marrow
3. Typhoid Fever
- S. typhi
- 1st week of infection: fever, malaise, aches, pains
- CONSTIPATION is the rule at this stage
- 2nd week: bacteremia
- ROSE-COLORED SPOTS on skin
- 3rd week of infection: Ab test
- best is serological test
TRASMISSION

- INVASINS:

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- Salmonellosis
- animal pathogen transferred to humans through
contaminated food (poultry and eggs)
- typhoid fever
- fecal-oral route
- contaminated H2O or food prepared by asymptomatic
Carriers

LABORATORY DIAGNOSIS
1. Culture and Isolation
-Specimen:
- 1st week: blood, bone marrow aspiration
- 2nd week: stool/ rectal swab
- 3rd week: urine
- chronic asymptomatic carriers lodge in
GALLBLADDER
- CM used for isolation
- XLD, BSA, EMB, SSA, SF.

- nonmotile
- non-lactose fermenters
- do not produce gas
-non-H2S production
-non-encapsulated
-non-spore former
- mannitol fermentation:
* non-mannitol fermenter:
- S. dysenteriae
* mannitol fermenter:
- S. sonnei
- S. boydii
- S. flexneri
- According to Somatic O Ag (A-D)
DETERMINANTS OF PATHOGENICITY
1. Invasion plasmid antigen
- attachment to and penetration of mucosal epithelium
but until submucosa only.
2. Intercellular spread proteins
- attachment to cytoskeleton proteins
- transfer of bacteria to adjacent cells not to tissues

2. Citrate Utilization (+)

3. TSIA (Alk/Alk) with H2S production
- some strain can produce glucose but all are non-lactose
Fermenters.

3.Shiga toxin
- inactivates 60s ribosomal subunit of mammalian cell
ribosomes > inhibit protein synthesis
- heat labile
- cytotoxic exotoxin

4. Phenylalanine Deaminase Test

CLINICAL DISEASE

5. Serotyping with use of anti-sera

1. Bacillary Dysentery
- bloody stool
- severe abdominal pain, frequently painful passage of
low volume stools with blood and mucus
- infection limited to colonic mucosa and submucosa

6. Typhoid Test
- ELISA
- High sensitivity with relatively low specificity
- affected by: chronic infections (false + rxn)
7. Widal Test
tither of 1:160
Vi carrier
O active infection
H past infection
PREVENTION AND CONTROL
- adequate sanitation
- immunization
- proper food preparation
- health education

SHIGELLA

2. Shigellosis
- no systemic manifestation
TRANSMISSION
- fecal-oral mode
- direct contact (increased sexual contact)
- contaminated food and water
LABORATORY DIAGNOSIS
1. cultural isolation
- SSA, EMB (colorless)
- stool and rectal swab
- S. dysenteriae
- pink to yellow

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- Colombia Agar

- lymphadenopathy (buboes) in the area drainage of the

flea bite

2. microscopic: gram (-) bacillus

3. Biochem: TSIA rxn (Alk/Alk)
a. IMVic Test: - + + - (Citrate difference to Salmonella)
b. Urease Prodn (-)
4. Methylene Blue Staining of Fecal Leukocytes
- mucosal invasion but non-specific
5. Sereny Test
- invasiveness of organisms
- A S drop of invasive strain and placed on the cornea of
guinea pig or rabbit
- (+) severe keratoconjunctivitis followed by ulceration
TREATMENT
- fluid replacement
- antibiotic average
- fluoroquinolones
CONTROL AND PREVENTION
- personal hygeine
- environmental sanitation.

YERSINIA
Y. pestis / Plague Bacillus
CHARACTERISTICS
- gram (-) coccobacillus with bipolar staining pattern
(SAFETY PIN)
- non-motile
- catalase (+)
- facultative anaerobe
- grows best at 28 degrees, colonies may take 2-5 days to
develop
- carried by a RAT FLEA known as Xenopsylla cheopis
DETERMINANTS OF PATHOGENICITY
- somatic Ag
1. F1: anti-phagocytic capsule (glycoprotein)
2. P1a: activate PLASMINOGEN, C3b and C5a

2. Pneumonic plague
- sever with mortality rate of 100%
- acquired by human to human transmission through
infected droplets
LABORATORY DIAGNOSIS
1. Gram Staining
2. Culture and Isolation
3. Broth Culture
- Stalactite streamers
- pellicle-like
4. Biochem
a. TSIA (Alk/Acid)
b. IMVic: _- + - TREATMENT
- antibiotic coverage
- streptomycin
- chloramphenicol
- tetracycline

Y. enterolytica
VIRULENCE FACTOR
- adhesins and invasins
- anti-phagocytic
- enterotoxins: similar to ST of E. coli
CLINICAL DISEASE
- gastroenteritis
- reactive arthritis
- ankylosing spindylitis
- nausea
- vomiting
RESERVOIRS:
-domestic animal, rodents
- isolated also in lakes and well waters
TRANSMISSION
- contaminated food and water
TREATMENT
- streptomycin
- chloramphenicol
- tetracycline

CLINICAL DISEASE
2 forms:
1. Bubonic plague

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