SHIGELLA Bacillary dysentery

BIOLOGICAL CHARACTERS

Gram-negative,
nonmotile,
facultatively anaerobic,
non-spore-forming rods
failure to ferment lactose
closely related with E coli
CLASSIFICATION
Four serogroups with multiple serotypes
Mannitol fermentation reaction distinguishes subgroup A(mannitol negative) from subgroups B,C and D (All are mannitol positive)

A (Sh. dysenteriae, 15 serotypes)

 Serotype 1- Bacillus originally described by Shiga (Sh. shiga) always catalase negative
 Serotype 2 – Sh. schmitzi forms Indole
 Serotype 3-7- Large-Sachs group

B (Sh. flexneri, 6 serotypes, further divided into subtypes, 2 antigenic variants)

 Serotype 6 occurs in 3 biotypes

C (Sh. boydii, 19 serotypes)

 Resembles S. flexneri biochemically but not antigenically
 Serotype13 reclassified as Esch. albertii
D (Sh. sonnei, 1 serotype)
 Antigenically homogenous, occurs in two forms- Phase I and Phase II
 Classified into many colicin types
 Most common Shigellosis in advanced countries
SHIGELLOSIS generic term for disease
 Low infectious dose (102-104 CFU)
 Humans are only reservoir
 Transmission by fecal-oral route
 5F’s- fingers, fluid, fomites, food, flies

 Incubation period = 1-3 days

 Watery diarrhea with fever; changing to dysentery
 Major cause of bacillary dysentery
 Outbreaks in daycare centers, nurseries, institutions
Dysentery = inflammation of intestines (especially the colon (colitis) of
the large intestine) with accompanying severe abdominal cramps,
tenesmus (straining to defecate), and frequent, low-volume stools
containing blood, mucus, and fecal leukocytes (PMN’s)
MANIFESTATIONS
Shigellosis
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
PATHOGENICITY
BACILLARY DYSENTERY--SHIGELLOSIS
THE EVENTS OF SHIGELLOSIS

Figure 20.15
VIRULENCE
1. Endotoxin
Released after cytolysis
cause fever, shock, bloody, mucoid stools, and abdominal
pain (cramps and tenesmus)

2. Exotoxin – Sh. dysenteriae type 1

 Enterotoxic- induce fluid accumulation
- ShET-1and 2
 Neurotoxic- damage endothelial cells of small blood vessels of
CNS resulting in polyneuritis, meningism and coma
VIRULENCE

3. Verocytotoxin (VT)
Sh. dysenteriae type 1; also by certain strains of E.coli
Cytotoxin acting on Vero cells
Chromosomally encoded
VT1 and VT2
2 subunits
 B – binds cytotoxin to cells
 A- inhibits protein synthesis (inactivates host cell 60S ribosome)
VIRULENCE

4. Invasive property
encoded by large plasmids – outer membrane protein
responsible for cell penetration called ‘virulence marker
antigens’ (VMA).
induce the endocytic uptake of Shigellae by M cells,
epithelial cells, and macrophages
deform the plasma membrane of contiguous cells
result in intercellular bacterial spread
Detection of VMA by ELISA serves as virulence test for
Shigellae
PATHOGENESIS
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
Preferentially attach to and invade into M cells in Peyer’s patches
(lymphoid tissue, i.e., lymphatic system) of small intestine
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
Histopathology of acute colitis following peroral infection with shigellae
COMPLICATIONS
Most often seen with Sh. dysenteriae type 1
Arthritis, toxic neuritis, conjunctivitis, parotitis, intussusception
(children)
Hemolytic uremic syndrome (HUS)
Reiter’s syndrome
LAB DIAGNOSIS
DIAGNOSIS -- SAMPLING

•Positive cultures are most often obtained from bloody mucus in

freshly passed stool specimens obtained during the acute phase of
disease
•Rectal swabs may also be used if the specimen is deposited in a
buffered glycerol saline holding solution
•Ideal specimen is a direct swab of an ulcer taken under
sigmoidoscopic examination
DIAGNOSIS --
Transport medium
Sach’s buffered glycerolsaline
Gram-negative broth
Enrichment medium
Selenite F broth (0.4%)
Salmonella Shigella broth
Diagnosis – Microscopy

Feature. Bacillary Amoebic.

Pus cells Numerous Scanty.

RBC Discrete. In clumps.

Macrophages With ingested Scanty.

RBC.

Eosinophils Absent. Present.

Charcot Layden Absent Present.

crystals.
E. histolytica Absent. Trophozoites.

Bacteria Non motile Plenty motile

.scanty.
 DIAGNOSIS -- ISOLATION

Primary differential/selective
media:
•MacConkey Agar
•Deoxycholate citrate agar (DCA)
•Xylose Lysine Deoxycholate colorless, non-lactose-fermenting colonies
agar (XLD)
•Salmonella-Shigella (SS) Agar
(contain bile salts& pH
indicators),
•Hektoen enteric (HE) agar…
DIAGNOSIS -- IDENTIFICATION

•Colony morphology
•MacConkey agar, DCA- Colorless colonies (NLF) except Sh. sonnei- pink
colonies due to late lactose fermentation
•XLD- red colonies without black center
•SS agar-colorless (NLF) with no blackening
• HE agar-green with fading to the periphery
•Staining- GNB
•Biochemical reactions
•Non-motile
•Urease, citrate, H2S and KCN negative
DIAGNOSIS --
•Slide agglutination tests with antisera
•Colicin typing – done for subgroup D (Sh. sonnei) strains
•Enzyme-linked immunosorbent assay (ELISA)
TREATMENT
1. Uncomplicated-
 self limiting
 Fluid supplement

2. Serious infections-
 Nalidixic acid/Norfloxacin/Fluoroquinolones
 R factor conferring antibiotic resistance to many drugs makes antibiotic therapy futile
PREVENTION
No vaccines
Improvement of environmental sanitation
Water supply
Sewage maintenance