ENTEROBACTERIACEAE

• a.k.a Enterics
The O Ag forms the outer part of the
General Characteristics
cell membrane.
4 major features:
The K Ag wraps around the cell like a
1. All ferment glucose.
capsule.
2. All reduce nitrates to nitrites.
The arms of the H Ag become wavy
3. None produce cytochrome oxidase.
flagella.
4. All are motile with flagellar arrangement as
peritrichous.
 Clinical Significance
• Non-motile enterics: * Most members of the Family Enterobacteriaceae are
1. Klebsiella present in the intestinal tract of animals and humans as
2. Shigella COMMENSAL FLORA.
3. Yersinia
Microscopic & Colonial Morphology
- gram negative
- non spore-forming
- facultative anaerobe bacilli

• Culture Media
non-selective media:
1. Sheep’s blood agar have little value • 2 Categories of Enterobacteriaceae based on the
2. Chocolate agar clinical infections they produce:
in ID
1. Opportunistic pathogens
differential & selective media: - for presumptive - part of the normal intestinal flora
identification of both humans and animals
1. MAC - [outside their habitat] è may
2. HE agar / Hektoen – highly selective produce serious extra-intestinal
3. XLD agar/Xylose-Lysine-Desoxycholate opportunistic infections
HE & XLD – contain Na thiosulfate & Fe+3 ammonium citrate, 1. Primary intestinal pathogens
which produce blackening of H2S- producing colonies - E.g. Salmonella, Shigella,
Yersinia enterocolitica
• Classification - considered true pathogens
- Ewing (1963) – proposed the use of tribes in - not commensal flora in the
classifying the members in the family gastrointestinal tract of humans
Enterobacteriaceae - produce infections that result from
- Edwards & Ewing’s Identification of the ingestion of contaminated food
Enterobacteriaceae and water.
- subsequent editions
OPPORTUNISTIC PATHOGENS
• Virulence & Antigenic Factors I. Escherichia coli
Factors that control virulence of Enterobacteriaceae: - gram (-) bacillus
1. the ability to colonize - common isolate from the colon flora
2. the ability to adhere - lactose (+) = dry, pink colony
3. the ability to produce various toxins - β-hemolytic colony on BAP
4. the ability to invade tissues - most are motile
Plasmids – mediate resistance to antimicrobials - possess both sex pili and adhesive fimbriae
• Antigens Used in Identifying different serologic - possess O, H, & K antigens
groups: *E. coli O groups – show cross-reactivity with similar antigens
1. O / Somatic antigen
- heat-stable Ag located in the cell wall Characteristics of E. coli:
2. H / Flagellar antigen 1. ferments glucose, lactose, trehalose and xylose
- heat-labile Ag found in the flagellum 2. indole & methyl red (+)
3. K / Capsular antigen 3. does not produce H2S, DNAse, urease, or
- heat-labile polysaccharide found in certain phenylalanine deaminase (PAD)
species such as the K1 Ag of E. coli & Vi Ag of 4. does not grow in the presence of KCN
Salmonella typhi. 5. cannot utilize citrate as a sole source of carbon
6. may be motile or non-motile
 7. (-) in Vogues-Proskauer test
• Clinical Infections
Theodore Escherich (1885)
- first described E. coli as a non-harmful
member of the colon flora
Gastrointestinal Infections
5 Major categories of diarrheogenic E. coli:
1. EPEC
2. ETEC
3. EIEC
4. EHEC
5. EAEC / EAggEC
• 5 Major categories of diarrheogenic E. coli:
1. Enteropathogenic E. coli (EPEC)
- cause infantile diarrhea
- outbreaks occur in hospital
nurseries and day care centers
- s/s:
• low-grade fever
• malaise vomiting
• diarrhea
• stool contains mucus
2. Enterotoxigenic E. coli (ETEC)
- associated with diarrhea of infants and
adults
- “traveler’s diarrhea”
- acquired by travelers visiting 3rd world
countries
- acquired by consuming contaminated
food or water
Major contributing factors in the spread &
transmission of the disease:
a. Poor hygiene
b. Inadequate sources of drinking water
c. Lack of proper sanitation
2. Enterotoxigenic E. coli (ETEC)
- Toxins produced:
a. LT / Heat-labile toxin
- action is similar to Vibrio cholerae toxin
- 2 fragments make up the LT:
B fragment – binds on the receptor site present on the
GM1ganglioside of the intestinal mucosa
 the binding facilitates the entry of the A fragment
A fragment – acts on adenyl cyclase
- activates conversion of ATP to cyclic
adenosine monophosphate (cAMP)
 the accumulation of cAMP in the intestinal mucosa
initiates the hypersecretion of electrolytes and
fluids into the lumen, resulting in a watery
diarrhea.
b. ST / Heat-stable toxin
- stimulates guanylate cyclase, causing the
increased producion of cyclic guanosine
monophosphate (cGAMP) and subsequent
hypersecretion
3. Enteroinvasive E. coli (EIEC)
- produce dysentery
- similar to that produced by Shigella
- occur in adults & children
- s/s:
• fever
• severe abdominal cramps
• malaise
• watery diarrhea accompanied by toxemia
• scanty stool containing pus, mucus and
blood
- non-motile
- do not ferment lactose
- cross-reactions between Shigella
- O Ag is seen
Sereny Test
- determines the organisms’ ability to
produce keratoconjunctivitis in the guinea pig
- one of the assays previously used to
determine the virulence of both Shigella and EIEC
4. Enterohemorrhagic (EHEC) O157:H7
- associated with hemorrhagic diarrhea,
colitis, and hemolytic uremic syndrome (HUS)
- low platelet count hemolytic anemia
kidney failure
- s/s:
• watery diarrhea bloody diarrhea
• crampy abdominal pain
• low-grade fever / no fever at all stool
contains no leukocytes
- infection is spread by undercooked
hamburger served at fast food restaurants,
unpasteurized milk and apple cider
- 2 cytotoxins:
a. Verotoxin I
b. Verotoxin II
verotoxin I
- a phage-encoded cytotoxin identical to the Shiga
toxin produced by Shigella dysenteriae type 1
- shows damage on vero cells (African green monkey
kidney cells)
- neutralized by the Ab against Shiga toxin
verotoxin II
- not neutralized by Ab to Shiga toxin
Culture media used:
a. MAC – contains sorbitol (SMAC)
- facilitates primary screening of E.
coli O157:H7
b. MUG assay (4-methylumbelliferyl-β-D-
glucuronide)
- a biochemical test that may be
used to screen for O157:H7
5. Enteroadherent E. coli (EAEC) / K. pneumoniae
Enteroaggregative E. coli (EAggEC)
- cause diarrhea by adhering to the
mucosal surface of the intestine
- s/s: Klebsiella pneumoniae
• watery diarrhea rod prokaryote (bacterium)
• vomiting
• abdominal pain
• dehydration
III. Enterobacter
OTHER E. COLI INFECTIONS - motile
a. UTI - (+) in Simmon’s citrate
o adhesins – mediates the adherence of urethrogenic - (+) in KCN broth
E. coli to the epithelial cells lining the urinary tract - methyl red (-)
o hemolysins – kill leukocytes and inhibit phagocytosis - Voges-Proskauer test (+)
and chemotaxis - produce ornithine decarboxylase
o aerobactin – an extracellular iron chelator a. Enterobacter cloacae
b. Septicemia & Meningitis b. Enterobacter aerogenes
- newborns acquire the infection in the
birth canal before or during delivery when the IV. Serratia
mother’s vagina is heavily colonized - opportunistic pathogens associated with
- infection also result if contamination of nosocomial outbreaks
the amniotic fluid takes place - ferment lactose slowly
- K1 Ag – the most documented - resistant to a wide range of antimicrobials
virulence-associated factor in neonatal - produce pigment
meningeal infections
E. coli a. Serratia marcescens produce a characteristic
b. Serratia rubidaea “prodigiosin” (pink to red pigment)

c. Serratia odorifera – gives off a dirty, musty odor like

potatoes
E. coli on the S. marcescens
human skin
surface

Serratia marcescens
Gram-negative prokaryote (bacterium)
II. Klebsiella
- found in the gastrointestinal tracts of
humans and animals
- large and very mucoid colonies
V. Hafnia
a. Klebsiella pneumoniae
- not known to cause gastroenteritis
- most common species
- possess a delayed (+) citrate reaction
- a frequent cause of lower respiratory tract
a. Hafnia alvei – the
infections
only species that belong to genus Hafnia
- also cause UTI and bacteremia
b. Klebsiella oxytoca
VI. Proteus – “Dienes Phenomenon”
- identical to K. pneumoniae
a. Proteus mirabilis
- indole (+)
b. Proteus vulgaris
c. Klebsiella ozaenae
- widely recognized human pathogens
- isolated from nasal secretions and cerebral
- isolated from urine, wounds, &
abscesses
- ear and bacteremic infections
d. Klebsiella rhinoscleroma
- produce swarming colonies on non-
- isolated from patients with rhinoscleroma
selective media, such as sheep’s blood agar
infection of the nasal cavity
- colonies produce “burned chocolate
- odor”
- produce H2S
- hydrolyze urea
*P. mirabilis is differentiated from P. vulgaris by the indole Erwinia – grows poorly at 37 0C
and ornithine decarboxylase tests: - fails to grow on selective media such as
P. mirabilis –indole (-), ornithine (+) EMB agar, MAC agar, and other differential
P. vulgaris – indole (+), ornithine (-) media used for the isolation of enterics.
P. vulgaris – sucrose (+)
- gives an acid/acid reaction in triple sugar iron (TSI) agar IX. Edwardsiella
- (-) for urea
c. P. penneri – formerly a P. vulgaris strain - (+) for lysine decarboxylase
- a newly recognized species that also - (+) for H2S
swarms on nonselective media - (+) for indole
d. P. myxofaciens – a species that has been isolated - (-) for Simmon’s citrate
only from gypsy moths a. Edwardsiella tarda – opportunistic
- characterized by the large amount of slime - cause bacteremia & wound infections
it produces b. Edwardsiella ictaluri – cause enteric septicemia in
fish
P. mirabilis c. Edwardsiella hoshinae – isolated from snakes, birds,
and water
Proteus mirabilis
Gram-negative, enteric, rod IX. Citrobacter
prokaryote - (+) methyl red test
(bacterium). - (+) Simmon’s citrate
- mistaken as Salmonella
P. vulgaris
a. Citrobacter freundii – associated with:
a. UTI
Proteus vulgaris b. Pneumonia
Gram-negative, rod prokaryote c. Intra-abdominal abscesses
(bacterium) d. Infectious diseases acquired in hospital
setting

VII. Morganella C. freundii

- has only 1 species:
a.Morganella morganii Citrobacter freundii
- formerly known as P. morganii Gram-negative, anaerobic
- has not been implicated in diarrheal illness prokaryote
- its role as an etiologic agent of diarrheal disease
remains to be further examined
- however, it is a documented cause of UTI
PRIMARY INTESTINAL PATHOGENS
VIII. Providencia
I. Salmonella
- consists of 4 species:
- gram (-)
a. P. alcalifaciens
- facultative anaerobe
- found in the feces of children with diarrhea
Biochemical features:
b. P. stuartii
1. non-lactose fermenters on TSI
- nosocomial outbreaks in burn units
2. (-) for indole, Voges-Proskauer, Phenylalanine,
- isolated from urine cultures
urease
c. P. rettgeri (formerly P. rettgeri)
3. (+) H2S on TSI
- documented pathogen of UTI
4. (-) on KCN
- has caused occasional nosocomial
outbreaks
I. Salmonella
d. P. rustigianii
a. Salmonella enteritidis
- rarely isolated
b. Salmonella choleraesuis
c. Salmonella typhi
X. ERWINIA & PECTOBACTERIUM
Virulence factors:
- both are plant pathogens
1. fimbriae – participates in the adherence to initiate
intestinal infections
2. ability to traverse intestinal mucosa
3. enterotoxin – cause gastroenteritis
 - a febrile disease that results from the
I. Salmonella ingestion of food contaminated with
Antigenic Structures: organisms originating from infected
1. somatic O Ag primary antigenic individuals
2. flagellar H Ag structures - humans = the only source of infection

capsular K surface Ag (Vi Ag) 3. Salmonella Bacteremia

- few strains may possess this - Salmonellosis – any disease caused by Salmonella
4. Carrier state
3. LPS (Lipopolysaccharides) – heat-stable O - gall bladder – the site of chronic carriage
Ag antimicrobial therapy – recommended if gall bladder
infection is not evident
I. Salmonella - cholecystectomy – the only solution to the chronic
Test: Widal test state of enteric carriers
O = active infection by S. typhi
H = exposed in endemic areas
= previous vaccination
= recovery stage
A = Paratyphi A infection
B = Paratyphi B infection

The salmon cruising in the bloodstream to infect the brain,

lungs, or bone. This systemic dissemination is usually
caused by Salmonella choleraesuis and does not involve the
GI tract.

Some people recovering from

Clinical Infections typhoid fever become chronic
1. Acute gastroenteritis / Food poisoning carriers, harboring Salmonella
- from ingestion of the organisms through typhi in their gallbladders and
contaminated food excreting the bacteria constantly.
- sources of infection:
a. poultry
b. milk
c. eggs and egg products S. typhi
d. Insufficiently cooked eggs
e. Domestic fowl
- chicken Salmonella typhi
- turkey rod prokaryote (bacterium)
- duck
f. Inadequate refrigeration of foods

Many animals can carry Salmonella. In

the U.S. there was even an epidemic of
salmonellosis from pet turtles.
Today in the U.S., Salmonella is most
commonly acquired from eating
chickens and uncooked eggs.
2. Typhoid fever
- clinical features:
• prolonged fever bacteremia
involvement of the RES
• dissemination to multiple organs
- caused by Salmonella typhi
II. Shigella
- cause bacillary dysentery
- named after Kiyoshi Shiga – Japanese
microbiologist (1896)
Dysentery – characterized by the presence of blood, mucus,
and pus in the stool
a. Shigella dysenteriae – Group A
b. Shigella flexneri – Group B
c. Shigella boydii – Group C
d. Shigella sonnei – Group D
 II. Shigella
general characteristics: Pneumonic form
a. non-motile - occurs secondary to bubonic plague when
b. do not produce gas from glucose organisms proliferate in the blood stream and
c. do not hydrolyze urea respiratory tract
d. (-) H2S on TSI
b. Yersinia enterocolitica
Shigella sonnei – slowly ferments lactose (pink on MAC) - (+) - the most common isolated species of
ONPG Yersinia
- from domestic swines, cats and dogs
II. Shigella - contact from household pets
Clinical Infections: - other sources:
“Gay Bowel Syndrome” • market meat
- in homosexuals • vacuum-packed beef
- caused by Shigella flexneri - survives in cold temperature

Humans – only known reservoir Infections manifest in several forms:

a. acute enteritis – the most common form of infection
MOT: b. Acute appendicitis-like syndrome
direct person-to-person contact c. Arthritis – common extraintestinal form of Y.
fecal-oral route -> source: carriers enterocolitica infection
d. Erythema nodosum – an inflammatory reaction
Shigellosis – any disease caused by Shigella characterized by red nodules

S. dysenteriae Culture medium:

CIN = Cefsulodin, irgasan, novobiocin

Shigella dysenteriae 3. Yersinia pseudotuberculosis

rod prokaryote (bacterium) - pathogen of rodents and guinea pigs in
particular
- reservoirs:
• turkey
S. sonnei • pigeons
• doves
• birds
Shigella sonnei • canaries
rod prokaryote (bacterium) - causes a disease characterized by caseous
swellings called pseudotubercules

Y. pestis
III. Yersinia
a. Yersinia pestis
- “bipolar staining” -> “safety pin Yersinia pestis
appearance” rod prokaryote (bacterium)
- gram (-), short, plump rod
- causes black plague / black death

plague – a life-threatening disease of rodents transmitted to

humans by fleas
- 2 forms of plague [in humans]
a. Bubonic /
Glandular form
b. Pneumonic form

Bubonic plague
- results from bites of an infected insect vector
- symptoms appear 2-5 days after infection