Dr.

Maha AdelMicrobiolology lecture-1-

I. Enterobacteriaceae (enteric = intestine) “Enterics”

 Enterics are ubiquitous in nature
 Except for few, most are present in the intestinal tract of animals and
humans as commensal flora; therefore, they are sometimes call “fecal
coliforms”
 Some live in water, soil and sewage

Family Enterobacteriaceae often referred to as “enterics”:

 Gram negative bacilli or coccobacilli
 Non-spore forming
 Colony morphology on BAP of little value, as they look the same,
except for Klebsiella
 Selective and differential media are used for initial colony evaluation
(ex. MacConkey, HE, XLD agars)
Four major features:

All ferment glucose (dextrose)

All reduce nitrates to nitrites ( NO3 to NO2 or all the way to N2 )

All are oxidase negative

All except Klebsiella, Shigella and Yersinia are motile by peritrichous

flagella

II. Pseudomanas- NOT Enterobacteriaceae

Gram-negative rods
oxidase-positive (cytochrome c oxidase; part of the electron transport chain)
respire only; not fermentative
polar flagella
(may produce a green water soluble pigment)
(colonies often develop a distinct sheen with age)

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 Dr.Maha AdelMicrobiolology lecture-1-

(ubiquitious; P. aeruginosa is major cause of nosocomial infections, UTIs, wounds,

pneumonia).
EXOTOXIN ENDOTOXIN

1. Released from the cell before 1. Integral part of cell wall

or after lysis
2. Endotoxin is LPS; Lipid A is toxic
2. Protein
component
3. Heat labile 3. Heat stable

4. Antigenic and immunogenic 4. Antigenic; ??immunogenicity

5. Toxoids can be produced 5. Toxoids cannot be produced

6. Specific in effect on host
6. Many effects on host
7. Produced by gram-positive and
gram-negative organisms 7. Produced by gram-negative
organisms only

Structure of Lipopolysaccharide

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Representative genera that are Medically Important Enterobacteriaceae

E. coli (Escherichia)
Klebsiella spp.

Morganella spp.

Proteus spp.

Salmonella spp.

Shigella spp.

Enterobacter
Citrobacter
Serratia spp.
Yersinia spp.

Culture methods
blood agar
selective/differential medium
MacConkey

on Mac
lac+ --> coliform
E. coli
Klebsiella
Enterobacter

lac- ---> other enteric

Pseudomonas

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Biochemical test
Voges-proskauer - butanediol pathway of fermentation
Motility
Indole - breakdown of tryptophan to produce indole
Citrate - use of citrate as a sole carbon source
Glucose
Lactose
Urea
TSI - glucose fermentation; lactose and/or sucrose fermentation; gas production; H2S
production

Virulence and Antigenic Factors of Enterics

▓ Ability to colonize, adhere, produce various toxins and invade tissues

▓ Some possess plasmids that may mediate resistance to antibiotics

▓ Many enterics possess antigens that can be used to identify groups

- O antigen – somatic, heat-stable antigen located in the cell wall

- H antigen – flagellar, heat labile antigen

- K antigen – capsular, heat-labile antigen

Based on clinical infections produced, enterics are divided into two

categories:

Opportunistic pathogens – normally part of the usual intestinal

flora that may produce infection outside the intestine

Primary intestinal pathogens – Salmonella, Shigella, and Yersinia sp

Escherichia coli
Most significant species in the genus

Important potential pathogen in humans

Common isolate from colon flora

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Positive indole and methyl red tests

Simmons citrate negative

Voges-Proskauer test negative

Does NOT produce H2S

Usually motile

Dry, pink (lactose positive) colony with surrounding pink area on

MacConkey

●Infections

Diseases

- Bacteremia (most commonly isolated gram-negative rod) ,

primarily from a genitourinary tract infection or a gastrointestinal
source.

- Urinary tract infection (most common cause of bacterial UTIs);

limited to bladder (cystitis) or can spread to kidneys (pyelonephritis)
or prostate (prostatitis)

- Gastroenteritis At least five different pathogenic groups cause

gastroenteritis (EPEC, ETEC, EHEC, EIEC, EAEC); most cause
diseases in developing countries, although EHEC is an important
cause of hemorrhagic colitis (HC) and hemolytic uremic syndrome
(HUS)

- Neonatal meningitis (usually with strains carrying the K1

capsular antigen). E. coli is one of the most common causes of
septicemia and meningitis among neonates; acquired in the
birth canal before or during delivery

- Intraabdominal infections (associated with intestinal perforation)

Most infections are endogenous

Gastrointestinal Infections

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Enteropathogenic (EPEC) – primarily in infants and children;

outbreaks in hospital nurseries and day care centers; stool has
mucous but not blood.

Enterotoxigenic (ETEC) – “traveler’s diarrhea”; watery

diarrhea without blood; self-limiting; usually not identified

Enteroinvasive (EIEC) – produce dysentery with bowel

penetration, invasion and destruction of intestinal mucosa;
watery diarrhea with blood.

Enterohemorrhagic (EHEC serotype 0157:H7) – associated

with hemorrhagic diarrhea and hemolytic-uremic syndrome
(HUS), which includes low platelet count, hemolytic anemia,
and kidney failure; potentially fatal, especially in young
children; undercooked hamburger and unpasteurized milk
have spread the infection.

Enteroaggregative (EAEC) – cause diarrhea by adhering to

the mucosal surface of the intestine; watery diarrhea;
symptoms may persist for over two weeks

Klebsiella species
Usually found in GI tract

Four major species, K. pneumoniae is mostly commonly isolated species

Possesses a polysaccharide capsule, which protects against

phagocytosis and antibiotics AND makes the colonies moist and
mucoid

Has a distinctive “yeasty” odor

Frequent cause of nosocomial pneumonia

Enterobacter species
Comprised of 12 species; E. cloacae and E. aerogenes are most common

Isolated from wounds, urine, blood and CSF

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Major characteristics

Colonies resemble Klebsiella

Motile

MR negative; VP positive

Serratia species
Seven species, but S. marcescens is the only one clinically important

Frequently found in nosocomial infections of urinary or respiratory tracts

Implicated in bacteremic outbreaks in nurseries, cardiac surgery, and burn

units

Fairly resistant to antibiotics

Major characteristics

Ferments lactose slowly

Produce characteristic pink pigment, especially when cultures are

left at room temperature

Proteus
All are normal intestinal flora

Opportunistic pathogens

All are lactose negative

P. mirabilis and P. vulgaris are widely recognized human pathogens

Isolated from urine, wounds, and ear and bacteremic infections

Both produce swarming colonies on non-selective media and have a

distinctive “burned chocolate” odor

Both are strongly urease positive

It exhibits characteristic “swarming”

Salmonella

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 Dr.Maha AdelMicrobiolology lecture-1-

Enrichment broth:Tetrathionat, selenit broth

Selective media: MacConkey, SS agar, Hektoen Enteric Agar, Bismuth

sulfide agar

Human ( S.Typhi, S. paratyphi)

Citrate (-)

Vi antigen positive

Gas from glucose negative

Tolerant to acids in phagocytic vesicles •

Can survive in macrophages and spread from the intestine to other •

body sites (particularly true of S. typhi)

Endotoxin •
Most infections are acquired by eating contaminated food products •

Direct fecal-oral spread in children •

S. typhi and S. paratyphi are strict human pathogens •

Individuals at risk for infection include those who eat improperly cooked •
poultry or eggs, patients with reduced gastric acid levels, and
immunocompromised patients

Infections occur worldwide, particularly in the warm months of the year •

Clinical Manifestations
- Gastroenteritis

- Enteric fever

- Bacteremia and vascular infections

- Carrier state

Asymptomatic colonization (primarily with S. typhi and S. paratyphi)

Enteric fever (also called typhoid fever [S. typhi] or paratyphoid fever [S.
paratyphi]).

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Incubation 5-21 days

Fever, relative bradycardia, leukopenia, anemia, constipation, rose spots

Neuropsychiatric manifestations

Mortality rate was 10-15% in preantibiotic era

Bacteremia (most commonly seen with S. typhi, S. paratyphi, S.

choleraesuis, and S. enteritidis, S.dublin)

Carrier state S.typhi %1-4, nontyphi Salmonella %0.2-0.6, no symptoms

Diagnosis

Culture

Stool, urine ,blood, rose spots, bone marrow

Selective media

Typhoid fever 1.week blood culture

≥3. week stool culture

Widal

Anti-O Ab

Anti-H Ab

Anti-Vi Ab (Long term carriers)

Diagnosis of typhoid fever

Blood cultures are positive during the first week and after the second week

Stool cultures and sometimes urine cultures are positive after the second
week.

The Widal test is a serological test for antibodies against Salmonella typhi.
One looks for a 4-fold rise in titer between acute and convalescent stages.

10% of those infected become short term carriers and a smaller % become
long-term carriers due to persistence of the bacteria in the gallbladder or
urinary bladder.

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 Dr.Maha AdelMicrobiolology lecture-1-

Shigella
S. dysenteriae, Shigella flexneri, Shigella boydii, and Shigella sonnei

S. sonnei is the most common cause of shigellosis in the industrial

world(developed contries).

S. flexneri is the most common cause in developing countries.

They are very much like Escherichia:

- Nonmotile

- Noncapsulated

- H2S negative

Endotoxin and genes for adherence, invasion, and intracellular •

replication

Permeability barrier of outer membrane •

Exotoxin (Shiga toxin) is produced by S. dysenteriae; disrupts protein

synthesis and produces endothelial damage

Hemolytic colitis (HC) and hemolytic uremic syndrome (HUS) associated

with Shigella

Humans are only reservoir for these bacteria

Disease spread person to person by fecal-oral route

Patients at highest risk for disease are young children in daycare centers,
nurseries, male homosexuals.

Relatively few organisms can produce disease (highly infectious).

Disease occurs worldwide with no seasonal incidence.

Gastroenteritis (shigellosis)

Most common form is an initial watery diarrhea progressing within 1 to 2

days to abdominal cramps and tenesmus (with or without bloody stools)

Asymptomatic carriage develops in a small number of patients (reservoir

for future infections)

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 Dr.Maha AdelMicrobiolology lecture-1-

A severe form of disease is caused by S. dysenteriae (bacterial

dysentery). Bloody diarrhae containing mucus,

Yersinia
Y. pestis, Yersinia enterocolitica, Yersinia pseudotuberculosis

Y. pestis is covered with a protein capsule, it causes systemic disease with

a high mortality, Capsule on Y. pestis is antiphagocytic

Y. pestis is also resistant to serum killing, Yersinia with genes for

adherence, cytotoxic activity, inhibition of phagocytic migration and
engulfment, and inhibition of platelet aggregation

Y. pestis is a zoonotic infection with humans the accidental host; natural

reservoirs include rats, squirrels, rabbits, and domestic animals

Disease is spread by flea bites or direct contact with infected tissues or

person to person by inhalation of infectious aerosols from a patient with
pulmonary disease.

Other Yersinia infections are spread through exposure to contaminated •

food products or blood products (Y. enterocolitica)

Y. pestis causes bubonic plague (most common) and pulmonary plague,

both having a high mortality rate

Other Yersinia species cause gastroenteritis (acute watery diarrhea or

chronic diarrhea) and transfusion-related sepsis

All Yersinia infections are zoonotic

Yersinia enterocolitica and Y. pseudotuberculosis –

clinical significance

Both are acquired by ingestion of contaminated food or

water.

Y. enterocolitica is a common cause of human disease,

whereas, Y. pseudotuberculosis is mainly a disease of
other animals.

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 Dr.Maha AdelMicrobiolology lecture-1-

Both cause a disease involving fever and abdominal

pain. Y. enterocolitica also causes a watery diarrhea.

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