Shigella

Introduction
• Cause bacillary dysentery (Shigellosis).

• Third most common cause of bacterial gastroenteritis in the United

States, after Campylobacter and Salmonella infection and ahead of E.
coli infection.

• Named after Kiyoshi Shiga, who isolated first member (Shigellae

dysenteriae) of this genus in 1896 from epidemic dysentery in Japan.
Classification
Shigella species are classified by four serogroups:

 Serogroup A: S. dysenteriae (12 serotypes)

(most common in industrial world)

 Serogroup B: S. flexneri (6 serotypes)

(most common in developing countries)

 Serogroup C: S. boydii (18 serotypes)

 Serogroup D: S. sonnei (1 serotype)

Morphology
• Gram negative short rods

• Size: 1-3µm x 0.5µm

• Non-motile, non-capsulated and non sporing.

Culture
• Aerobes and facultative anaerobes.

• Grows on ordinary medium at temperature range

of 10-400C ( optimum: 370C)

• Colonies are about 2mm in diameter, circular, convex, smooth and

translucent.
• On MacConkey agar (MA) and Deoxycholate citrate agar (DCA)
colonies are colourless (non-lactose fermenting- NLF)
• On xylose lysine deoxycholate (XLD) agar ; appear red without black
centres.
• On Salmonella-Shigella (SS) agar : colourless with no blackening.
Biochemical reaction:
• Ferments glucose with acid
• Do not ferments lactose except Sh. Sonnei (late lactose fermenter)
• Serogroup B, C and D ferments mannitol but not serogroup A
• H2S is not produced
• Do not Utilise citrate
• Catalase test is positive ( except Sh.dysenteriae type 1).
Toxins
• Endotoxin:
- release after autolysis,
- has irritating effect on intestinal wall causing diarrhoea and intestinal ulcers.

• Exotoxin:
acts as
- enterotoxin(Induces fluid accumulation) and
- neurotoxin (damages the endothelial cells of small blood vessels of the CNS
results in polyneuritis, coma and meningism).
• Verocytotoxin (VT):
- Acts on vero cells ( a cell line derived from African green monkey
kidney cell in 1977)
- Two verocytotoxins, VT1 and VT2 are recognized.
- VT comprises two subunits- A and B
- Subunit B binds the toxin to cells while subunit A inhibits protein
synthesis.
Pathogenesis
• Incubation period:
- usually less than 48 hours but varies between 1-7 days.

• Mode of transmission :
- faeco- oral route

• Minimum infective dose:

- as few as 10-100 bacilli
Pathogenesis
Two-stage disease:
 Early stage:
 Watery diarrhea attributed to the enterotoxic activity of Shiga
toxin following ingestion and noninvasive colonization,
multiplication, and production of enterotoxin in the small
intestine

 Fever attributed to neurotoxic activity of toxin

 Second stage:
 Adherence to and tissue invasion of large intestine with typical
symptoms of dysentery

 Cytotoxic activity of Shiga toxin increases severity

 Invasiveness in Shigella-Associated Dysentery
 Penetrate through mucosal surface of colon (colonic
mucosa) and invade and multiply in the colonic
epithelium but do not typically invade beyond the
epithelium into the lamina propria (thin layer of fibrous
connective tissue immediately beneath the surface epithelium of
mucous membranes)

 Preferentially attach to and invade into M cells in Peyer’s

patches (lymphoid tissue, i.e., lymphatic system) of small
intestine
 Invasiveness in Shigella-Associated Dysentery(cont.)
 M cells typically transport foreign antigens from the
intestine to underlying macrophages, but Shigella can
lyse the phagocytic vacuole (phagosome) and replicate
in the cytoplasm
 Note: This contrasts with Salmonella which multiplies in
the phagocytic vacuole

 Actin filaments propel the bacteria through the

cytoplasm and into adjacent epithelial cells with cell-
to-cell passage, thereby effectively avoiding antibody-
mediated humoral immunity (similar to Listeria
monocytogenes)
 Characteristics of Shiga Toxin
 Enterotoxic, neurotoxic and cytotoxic
 Encoded by chromosomal genes
 Two domain (A-5B) structure
 Similar to the Shiga-like toxin of enterohemorrhagic E.
coli (EHEC)
 Shiga Toxin Effects in Shigellosis

Enterotoxic Effect:
 Adheres to small intestine receptors

 Blocks absorption (uptake) of electrolytes, glucose,

and amino acids from the intestinal lumen
 Shiga Toxin Effects in Shigellosis (cont.)
Cytotoxic Effect:
 B subunit of Shiga toxin binds host cell glycolipid
 A domain is internalized via receptor-mediated endocytosis
(coated pits)
 Causes irreversible inactivation of the 60S ribosomal subunit,
thereby causing:
 Inhibition of protein synthesis
 Cell death
 Microvasculature damage to the intestine
 Hemorrhage (blood & fecal leukocytes in stool)

Neurotoxic Effect:
Fever, abdominal cramping are considered signs of neurotoxicity
Clinical syndrome:
• Dysentery characterized by frequent painful passage of low volume
stools containing blood, pus and mucous, with severe abdominal
cramps.

• Sh. Dysenteriae type 1 infection may cause

- Severe diarrhoea,
- Neurological complications (polyneuritis),
- Myocarditis or joint effusions as a result of toxaemia.
- Renal damage may lead to haemolytic uraemic nephritis.
Laboratory diagnosis:
• Sample: fresh stool, swab of an ulcer taken under sigmoidoscopic
examination.

• Microscopy:
- Saline and iodine preparation: shows large number of pus cells, erythrocytes
and marophages along with bacteria.

• Culture:
- Selenite-F broth- used as enrichment medium
- Subculture is done on MA, DCA, XDA, SS etc…
• Biochemical test:
- Urease, citrate and KCN negative.

- On TSI: K/A with gas and H2S negative

- Indole reactions are mixed, positive and negative, with the exception
of S. sonnei, which is always indole negative.

• Slide-agglutination test
Prevention and treatment
• Supportive measures: fluid replacement

• Antibiotics:
- Ampicillin, amoxycellin may be given for young patients.
- Nalidixic acid or norfloxacin in severe cases.
Thank you