‫‪SAWA UNIVERSITY‬‬ ‫عنوان المحاضرة‪:‬‬

‫‪Enterobacteriaceae‬‬
‫‪5. Shigella.‬‬

‫‪-‬‬ ‫‪6. Salmonella.‬‬

‫جامعة ساوة‬

‫كلية التقنيات الصحية والطبية‬

‫قسم تقنيات المختبرات الطبية‬

‫المرحلة‪ :‬الثانيــة‬

‫بكتــريــا طبيــة ‪2‬‬

‫‪@AHMEDFIRAS2005_BOT‬‬

‫اسم المحاضر‪ :‬الـدكتـور معيـه حسـه‬

‫رقم المحاضرة (المحاضرة الرابعــة)‬
 CONTENTS OF THIS TEMPLATE

● Introduction
● Classification of Shigella
● Virulence Factors
● Pathogenesis of Shigella
● Laboratory Diagnosis
● Classification of Salmonella
● Virulence Factors
● Pathogenesis of Salmonella
● Laboratory Diagnosis
Genus: Shigella
 Familly: Enterobacteriaceae
 Tribe: Escherichia
 Genus: Shigella
 Discovered by Kiyoshi Shiga in 1898.
 It is the causative agent of human shigellosis.

 Classification:
 There are more than 40 serotypes.
 The classification of shigellae relies on biochemical and antigenic
characteristics (O antigens).
 The pathogenic species are Shigella sonnei, Shigella flexneri,
S. dysenteriae, and Shigella boydii.
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 Important Properties:

 Shigellae are:
 Short Gram-negative rods.
 Non–lactose-fermenting.
 Resistant to bile salts
 Divided into four groups: A, B, C, and D according to (O) antigen.

 Shigella can be distinguished from salmonellae by three criteria:

1. They produce no gas from the fermentation of glucose.
2. They do not produce H2S.
3. They are non-motile.

4
 Virulence Factors:
1. K. capsular antigen
2. O. antigen (HL)
3. Shiga toxin: with cytotoxic and neurotoxic activity.

SHI6£LI.A

5
On MacConkey agar
 Pathogenesis of Shigella:

1. Efltry

2. Disease
Oehydra.tion
Vomiting
Diarrhea
Dysentery
Tenesmus
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3-. Exit
n 6
 Shigella causes bacillary dysentery.
 Low infective dose < 200 bacilli (can be transmitted easily unlike
salmonella).
 (More serious and virulent than salmonella).
 Incubation period= 1-3 days
 Upon ingestion, the bacteria pass through the gastrointestinal tract until
they reach the small intestine.
 There they begin to multiply until they reach the large intestine.
 In the large intestine, the bacteria cause cell injury and the beginning stages
of Shigellosis via two main mechanisms:
1. Direct invasion of epithelial cells in the large intestine and production of
enterotoxin 1 and enterotoxin 2.
2. High fever, chill, abdominal cramp and pain accompanied by tenesmus,
bloody stool with mucus & WBC and HUS are involved. 7
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 Laboratory Diagnosis:
 Specimens: include fresh stool, mucus flecks, and rectal swabs for culture.
 Large numbers of fecal leukocytes and some red blood cells often are seen
microscopically.
 Culture: The materials are streaked on differential media (e.g. MacConkey or EMB agar) and on
selective media (Hektoen enteric agar or xylose-lysine-deoxycholate agar (XLD), which
suppress other Enterobacteriaceae and Gram-positive organisms.

 Colorless (lactose-negative) colonies are inoculated into TSI agar.

 Organisms that fail to produce H2S, that produce acid but not gas in the butt and an
alkaline slant in TSI agar medium.

 Salmonella Shigella (SS) Agar: Shigella Clear, colorless, transparent.

 XLD- Agar: Shigella flexneri , Red Colonies.

 TSI-Agar: Salmonella Alkaline slant/acidic butt (K/A); H2S and Gas. 9
Genus: Salmonella
 Introduction:. The organisms are named after the American veterinary
pathologist Daniel Elmer Salmon in 1885.
 Currently, there are three recognized species: S. enterica, S. bongori and
S. subterranean .
 Salmonella is found worldwide in cold and warm-blooded animals
(including humans), and in the environment.

 They cause illnesses such as typhoid fever, paratyphoid fever, and
foodborne illness.

10
 Classification:
 The members of the genus Salmonella were originally classified on the basis of
epidemiology; host range; biochemical reactions; and structures of the O, H, and Vi
(when present) antigens.

 Salmonella spp. have both H and O antigens.

 There are over 60 different O antigens, and individual strains may possess several O
and H antigens; the latter can exist in variant forms, termed ‘phases’.

Salmonella serotype Typhi also has a capsular polysaccharide antigen referred to as

‘Vi’ (for virulence), which is related to invasiveness
 Over 2500 serotypes are distinguished, most of which belong to the species S. enterica .

 However, many these have been given binomial names (e.g.

of
Salmonella typhimurium and Salmonella enteritidis ), although they are not separate
species.
 In clinical practice, laboratories identify microorganisms according to their binomial
name. 11
 Important Properties: Salmonellae are motile rods that characteristically
ferment glucose and mannose without producing gas but do not ferment
lactose or sucrose.
 Most salmonellae produce H2S.
 They are often pathogenic for humans or animals when ingested.

 Virulence Factors:
1. Type III secretion systems: which facilitate secretion of virulence factors of
Salmonella into host cells.

2. Endotoxin: Endotoxin is responsible for many of the systemic

manifestations of the disease caused by Salmonella spp.

3. Fimbriae: The species-specific fimbriae mediate binding of Salmonella to M

(microfold) cells present in Peyer patches of the terminal part of the small intestine.

 These M cells typically transport foreign antigens, such as bacteria to the 12

underlying macrophages for clearance.
4. Acid tolerance response gene: The acid tolerance response (ATR) gene
protects Salmonella spp.
 from stomach acids and the acidic pH of the phagosome, thereby
facilitating survival of bacteria in phagosomes
5. Enzymes: Catalase and superoxide dismutase are the enzymes that protect
the bacteria from intracellular killing in macrophages.

 Pathogenesis of Salmonella:
 The three types of Salmonella infections (enterocolitis, enteric fevers, and
septicemia), Have different pathogenic features.

13
 1. Entry
3. Disease·
Gasl roenteritis -...---- 2. Spread
Diarrhea (infrequent, typhoid tever)

4. E:xit
(gallbladder-carrier state)

14
 1. Enterocolitis: is characterized by an invasion of the epithelial and
sub-epithelial tissue of the small and large intestines.
2. In typhoid and other enteric fevers, infection begins in the small intestine,
but few gastrointestinal symptoms occur.
3. Septicemia accounts for only about 5−10% of Salmonella infections and
occurs in one of two settings: a patient with an underlying chronic disease:
such as (sickle cell anemia or cancer), or a child with enterocolitis.
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 Laboratory Diagnosis:
 In enterocolitis: the organism is most easily isolated from a stool sample in:
 Selective media e.g. 1. XLD (Xylose lysine deoxycholate agar),
2. DCA (deoxycholate citrate agar), 3. salmonella-shigella (SS) agar. and

 Enrichment media, e.g. selenite broth; identification of Salmonella spp.

 by biochemical agglutination tests. Phage typing can be used for typing
individual strain.
 Salmonella Shigella (SS) Agar: salmonella colorless, transparent, with a
black center if H2S is produced
 XLD- Agar: Salmonella Typhi red Colonies, black centers.
 TSI-Agar: Salmonella Alkaline slant/acidic butt (K/A); + H2S and Gas +.

16
 In the enteric fevers: a blood culture is the procedure most likely to reveal
the organism during the first weeks of illness.
 Stool cultures may also be positive, especially in chronic carriers in whom
the organism is secreted in the bile into the intestinal tract.
 Urine culture results may be positive after the second week.

 Serologic Methods:
1. Agglutination test
2. Tube dilution agglutination test (Widal test):
 Serum agglutinins rise sharply during the second and third weeks of
S serotype Typhi infection.

17