Lec-10

Salmonella & Shigella

Prepared by:
Dr. Hanadi A.Jasim
Assist Professor
Apr. 11th 2023

Objective
 Distinguish the pathogenic members.
 Know the features and mechanism to cause
infections.
 Specify the infections that caused by these
bacteria.
 Describe the methods for lab. Diagnosis.

2
 Clinical case
 A 48-year-old man presents to the emergency room with
2 days of crampy abdominal pain, nausea, vomiting,
diarrhea, and fever. He has not had any blood in his stool.
He denies contact with anyone with similar symptoms
recently. He has not eaten any raw or unprocessed foods
recently. The only food that he did not prepare himself in
the past week was a breakfast of eggs “sunny-side up”. On
examination, he is tired appearing; his temperature is
37.7°C (99.9°F); A rectal exam reveals only heme-
negative watery stool. Gram stain revealed gram negative
rods.
◆ What is the most likely etiologic agent of this infection?

 Pathogenic for humans or

animals when acquired by
the oral route.
 Transmitted from animals
and animal products to
humans and cause
 Enterocolitis,
 Enteric fevers such as
typhoid fever,
 Septicemia
 Important Properties Salmonellae
 Gram-negative rods
 Do not ferment lactose
 Do produce H2S.
 Motile with peritrichous flagella
 Grow readily on simple media,
 Ferment glucose and mannose without producing gas
 They survive freezing in water for long periods.
 Salmonellae are resistant to certain chemicals (eg, brilliant
green, sodium tetrathionate, sodium deoxycholate) that
inhibit other enteric bacteria;

Antigenic structure;
 Are important for taxonomic and
epidemiologic purposes.
1. Cell wall O Ag
2. Flagellar H Ag
3. Capsular Vi (virulence)

 The O antigens, which are the outer

polysaccharides of the cell wall
 The Vi antigens (capsular polysaccharides)
antiphagocytic
 The Vi antigens are also used for the
serotyping of S. typhi in the clinical laboratory.
 Classification and naming
 There are three methods for naming the salmonellae.
1- Ewing divides the genus into three species:
S. typhi, Salmonella choleraesuis, and Salmonella
enteritidis.
2- Kaufman and White assign different species names to each
serotype;
 usually named for the city in which they were isolated.
Salmonella dublin
3- Based on DNA hybridization analysis.
 In this scheme, S. typhi is not a distinct species but is classified as
Salmonella enterica serotype (or serovar) typhi.
 All three of these naming systems are in current use.

 Clinically, the Salmonella species are often thought of

in two distinct categories, namely,
1. The typhoidal species (those that cause typhoid fever)
2. The nontyphoidal species (those that cause diarrhea
[enterocolitis] and metastatic infections, such as
osteomyelitis).
 The typhoidal species are S. typhi and S. paratyphi.
 The nontyphoidal species are the many serotypes of S.
enterica.
 Epidemiology
 Ingestion of food and water contaminated by human and
animal wastes.
 S. typhi, the cause of typhoid fever, is transmitted only
by humans,
 all other species have a significant animal as well as human
reservoir.
 Human sources are either:
1. Persons who temporarily excrete the organism
during or shortly after an attack of enterocolitis
2. Chronic carriers who excrete the organism for
years.
 The most frequent animal source is poultry and
eggs.

Virulence factors
 Pathogenesis & Clinical findings
 The three types of Salmonella infections have different pathogenic
features:
(1) Enterocolitis is characterized by:
 Invasion of the epithelial and subepithelial tissue of
the small and large intestines.
 Strains that do not invade do not cause disease.
 The organisms penetrate both through and between
the mucosal cells into the lamina propria, with
resulting inflammation and diarrhea.
 Bacteremia is infrequent
 Salmonella required at least 100,000 organisms.
 Clinical Findings Enterocolitis
 Incubation period of 12 to 48 hours,
 Nausea
 Vomiting
 Abdominal pain
 Diarrhea, which can vary from mild to severe, with or
without blood.
 Usually the disease lasts a few days, is self limited,
causes non bloody diarrhea, and does not require
medical care except in the very young and very old.
 Salmonella typhimurium is the most common
species.

Enterocolitis
 (2) Typhoid and other enteric fevers
 Infection begins in the small intestine.
 The organisms enter, multiply in the mononuclear
phagocytes of Peyer’s patches, and then spread to the
phagocytes of the liver, gallbladder, and spleen.
 Bacteremia,
 Survival and growth of the organism within phagosomes
 Invasion of the gallbladder, can result in establishment of
the carrier state and excretion of the bacteria in
the feces for long periods.

Clinical Findings Typhoid fever

 Incubation period of 10–14 days

 The onset of illness is slow,
 Fever and constipation rather than vomiting and
diarrhea predominating.
 Diarrhea may occur early
 Bacteremia
 Enlarged spleen occur.
 Rose spots are associated with typhoid fever but occur
only rarely.
 The disease begins to resolve by the third week,
 About 3% of typhoid fever patients become chronic
carriers.
 (3) Septicemia
 Accounts for only about 5% to 10% of Salmonella
infections
 occurs in one of two settings:
1. A patient with an underlying chronic disease, such
as sickle cell anemia or cancer,
2. A child with enterocolitis.

 Bacteremia results in the seeding of many organs,

with osteomyelitis, pneumonia, and meningitis as
the most common sequelae.
Laboratory Diagnosis Specimens
1- Blood for culture must be taken repeatedly.
 In enteric fevers and septicemias, blood culture results are
often positive in the first week of the disease.
 Urine culture results may be positive after the second
week.

2- Stool specimens also must be taken repeatedly.

 In enteric fevers, the stools yield positive results from the
second or third week on;
 in enterocolitis, the stools yield positive results during the
first week.
 A positive culture of biliary tract indicate carriers.
 Laboratory Diagnosis Bacteriology
(See lab- 15)
 A- Cultivation:
1. Differential medium cultures: EMB, MacConkey
2. Selective medium cultures: Salmonella-Shigella
(SS) agar,
3. Enrichment cultures: (specimen usually stool)
-Selenite F or Tetrathionate broth
B- Biochemical reaction (TSI test ) and (urease test)
C- Serologic Methods
1. Agglutination test
2. Tube dilution agglutination test (Widal test)

2. Tube dilution agglutination test (Widal test)

 Serum agglutinins rise sharply during the second and third
weeks of S. typhi and paratyphi infection.
 The Widal test to detect these antibodies against the O and H
antigens
 The titre of the patient serum using Widal test antigen
suspensions is the highest dilution of the serum sample that
gives a visible agglutination.
 O antigen of greater than 1:160 is considered positive(active inf.)
 H antigen of greater than 1:160 is considered positive(past inf. or
immunization)
 High titer of antibody to the Vi antigen occurs in some carriers.
 Alternatives to the Widal test include rapid colorimetric and EIA
methods.
 Mac agar
Widal test
XLD agar

SS agar TSI

Treatment Ceftriaxone or ciprofloxacin.

Prevention

 Public health and personal hygiene measures.

 Proper sewage treatment,
 A chlorinated water supply,
 Handwashing prior to food handling,
 Pasteurization of milk,
 Proper cooking of poultry, eggs, and meat
 Shigella
 Shigella species cause enterocolitis.
 Enterocolitis is often called bacillary dysentery.
 The term dysentery refers to bloody diarrhea.
 The natural habitat of shigellae is limited to the intestinal tracts
of humans and other primates, where they produce bacillary
dysentery.

The pathogenic species are

 Shigella sonnei,
 Shigella flexneri,
 Shigella dysenteriae,
 Shigella boydii
 Shigella Important Properties
 Shigellae are non–lactose-fermenting,
 Gram-negative rods that can be distinguished
from salmonellae by three criteria:
1. Produce no gas from the fermentation of
glucose,
2. Do not produce H2S,
3. Nonmotile
 All shigellae have O antigens (polysaccharide) in
their cell walls, and these antigens are used to divide
the genus into four groups: A, B, C, and D.

Virulence mechanisms

 Virulence mechanisms of Shigella include

 Ability to invade the intestinal mucosa
 Production of shiga toxin, which acts
to destroy the intestinal mucosa once the
organism has invaded the tissue.
 Pathogenesis & Epidemiology
 Shigellae are the most effective pathogens among the enteric
bacteria.
 Infection almost always limited to the gastrointestinal tract;
 Bloodstream invasion is quite rare.
 Shigellae are highly communicable;
 Ingestion of as few as 100 organisms causes disease,
 Shigellosis is only a human disease
 No animal reservoir.
 The organism is transmitted by the fecal–oral route.
 The four Fs—Fingers, Flies, Food, And Feces—are the
principal factors in transmission.

Pathogenesis & Epidemiology

 Outbreaks occur in day care nurseries and in mental hospitals,
 There is no prolonged carrier state
 Shigellae, produce bloody diarrhea (dysentery) by invading
the cells of the mucosa of the distal ileum and colon.
 Local inflammation & ulceration
 Although some strains produce an enterotoxin (called Shiga
toxin), invasion is the critical factor in pathogenesis.
 Shiga toxins very similar to those produced by Shigella are
produced by enterohemorrhagic E. coli O157:H7 strains that
cause enterocolitis and HUS.
 Clinical Findings
 Incubation period of 1 to 4 days,
 Fever & abdominal cramps, followed by diarrhea, which
may be watery at first but later contains blood and
mucus.
 The disease varies from mild to severe depending on two
major factors:
1. The species of Shigella
2. The age of the patient,
 Shigella dysenteriae, -------severe disease
 Shigella sonnei, -------------mild disease,
 The diarrhea frequently resolves in 2 or 3 days;
 Laboratory Diagnosis (See lab-15)
A. Specimens
 fresh stool, mucus flecks, and rectal swabs for culture.
 Large numbers of fecal leukocytes and some red blood cells often
are seen microscopically.
 B. Culture
 MacConkey or EMB agar
 Hektoen enteric agar or XLD agar
C. Biochemical: TSI agar medium, Motility test.
D. Slide agglutination by specific Shigella antisera..
E. Nucleic Acid Amplification Tests

Treatment
 Fluid and electrolyte replacement.
 In mild cases, no antibiotics are indicated.
 Ciprofloxacin is the drug of choice,

Prevention
 Sewage disposal,
 Chlorination of water, and
 Personal hygiene (handwashing by food handlers).
 There is no vaccine
 In which of the following sites is S. typhi most
likely to be found during the carrier state?
A. Blood
B. Gallbladder
C. Kidney
D. Liver
E. Spleen

 A 4-year-old has fever and diarrhea. Blood

culture grows a gram negative rod. This is
most likely to be which of the following?
A. Group B Streptococcus
B. Listeria species
C. Salmonella species
D. Shigella species
Thanks