Campylobacter

Dr. Sundus Javed

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 History
• From greek curved rods
• Helicobacter previously classified as campylobacter
• Discovered in1886 by Theodor Escherich from an infant who died of
cholera and called the disease “cholera infantum.”
• 1913, McFayden and Stockman identified from an aborted sheep-
Vibrio fetus (now known as Campylobacter fetus).
• In 1972, Dekyser and Butzler isolated a Campylobacter strain from
blood and feces of a woman who suffered from hemorrhagic enteritis.
Considered a zoonotic pathogen.
• Campylobacter-related infection is the highest among all the
foodborne bacterial infections in the US (1.9 million cases per year)
• Many cases are associated with consumption of chicken.
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 Characteristics
• Campylobacteracae family consists of two genera Campylobacter and
Arcobacter
• Campylobacter species are Gram-negative, nonspore-forming,
curved, S-shaped, or spiral helical rods with approximately 0.5–5.0
μm in length.
• Single polar flagellum at one or both ends
• Exhibit corkscrew-like motion.
• In older cultures, bacteria may actually appear as spherical or coccoid
bodies which is a dormant, viable but nonculturable state (VBNC)

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 Characteristics
• They are fastidious curved rods with stringent growth requirements.
Campylobacter is microaerophilic, and several of the species are
thermophilic and are unable to grow below 30 °C.
• Arcobacter are aerotolerant and can grow below 30 °C.
• Both Campylobacter and Arcobacter are routinely isolated from
livestock, poultry, and water.
• Outbreak of Campylobacter is associated with meat, poultry, and milk.
• Of 16 species of Campylobacter, C. jejuni is responsible for 95% of
the outbreaks and is considered the most predominant pathogen.

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Campylobacter Classification based on Biochem
characteristics

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Campylobacter Classification based on Biochem
characteristics

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 Pathogenesis
• Campylobacter pathogenesis depends on the expression of several
virulence factors that control motility, chemotaxis, quorum sensing,
bile resistance, adhesion, invasion, toxin production, growth inside
cells, and iron acquisition.
• Bacteria possibly induce their own internalization through signaling
events and rearrangement of host cytoskeletal structure, and survival
inside the epithelial cells by expressing superoxide dismutase and
catalase to deactivate host oxidative stress defense.
• Cytolethal distending toxin (CDT) arrest cell cycle division and
disrupts the absorptive function of villous epithelial cells and promote
diarrhea.

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 Campylobacter associated host cell damage

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 Pathogenesis
• Campylobacter may cause fatal infection in immunocompromised
patients.
• Patients suffering from C. jejuni infection may also develop Guillain-
Barré syndrome characterized by generalized paralysis and muscle
pain and the Reiter’s syndrome characterized by arthritis in knee
joints or lower back.
• The pathogenic mechanism of Arcobacter is unknown, but they cause
diarrhea in humans (in children) and abortion and stillbirth in cows,
sheep, and pigs..

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 Pathogenesis
• In association with food or water, campylobacters enter the host
intestine via the stomach acid barrier
• colonize the distal ileum and colon.
• adhesion to intestinal cell surfaces
• perturb the normal absorptive capacity of the intestine directly by
damaging epithelial cell function, by cell invasion or the production of
toxin(s),
• or indirectly, by the initiation of an inflammatory response.
• any combination may have a role depending on the host status and
attributes of the infecting strain.

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 Chemotaxis and motility
• campylobacters have mechanisms to detect chemical
• gradients and linked motility functions that enable the
• cell to move up or down the gradient.
• mutagenized, non-chemotactic mutants fail to colonize intestine of
suckling mouse intestine.
• various chemoattractants, including mucin, L-serine and L-fucose, and
several bile acids have chemorepellant effects.
• cheY, does not affect motility or invasion but does result in a loss of
chemotaxis in vitro.

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 Chemotaxis and motility
• flagellum and cell shape give campylobacters an unusually high level
of motility in viscous environment
• The unsheathed flagellum exhibits antigenic variation.
• The locus has been mapped and, in the majority of isolates, contains
two adjacent genes, flaA and flaB.
• the absence of FlaA, the flaB gene encodes a flagellin protein that
forms a short, truncated non-functional flagellum. FlaA is essential for
colonization
• FlaB is incorporated into the whole filament in small amounts and flaB
mutant produces a normal length flagellum with slightly decreased
motility in comparison to the wild-type strain. Defined mutants have

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 Attachment and Invasion
• C. coli, C. concisus, C. fetus, C. rectus and C. upsaliensis all have the
ability to invade host cells
• C. coli strains frequently harbor cadF (mediates binding to fibronectin,
aids attachment to host cells), ciaB and pldA genes (encode invasion-
associated proteins).
• C. lari and C. upsaliensis harbor genes for the cell binding protein
PEB1 and a putative fibronectin-binding protein.
• attach and invade T84, Caco-2, and HeLa cell and primary intestinal
cells, a process that is also dependent on host microfilaments and/or
microtubules.
• adhere to small intestinal mucin and bind to human host lipid
receptors.
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 Attachment and Invasion
• C. concisus preferentially attach to the intercellular epithelial junction,
invade intestinal epithelial cells and generate a membrane-ruffling
effect on entry in a process mediated by the polar flagellum and
possibly other bacterial surface proteins
• Attachment to the intestinal epithelium increases host barrier
permeability, decreases transepithelial electrical resistance, and
results in a loss of tight junction proteins.
• both oral and fecal-affiliated C. concisus strains initiate apoptotic
events in mucus-producing colonic epithelial monolayers an
observation that coincided with compromise barrier functions.
• presence of a fibronectin-binding protein in the outer membrane of the
bacteria, which facilitate bacterial attachment to host cells.
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 Attachment and Invasion
• Binding of C. fetus to primary human colonic intestinal cells is
enhanced by extracellular matrix components of the host cell, such as
fibronectin, and to some extent, type I collagen and laminin, but not
type IV collagen and heparan sulfate.
• Invasion and translocation across a Caco-2 monolayer by C. fetus
has also been observed. C. hominis, C. showae and C. ureolyticus
attach to, but do not seem to be able to invade, Caco-2 intestinal
epithelial cells, a finding that suggests that not all members of the
Campylobacter genus are invasive.
• During invasion, C. jejuni is encapsulated within a vacuole and avoids
delivery to lysosomal compartments, thereby circumventing
antimicrobial agents and prolonging intracellular survival.
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 C. jejuni invasion into epithelial cells

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 clinical outcomes of infection

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 Cytolethal distending toxin
• cytolethal distending toxin (CDT) is the best characterized toxin
associated with Campylobacter
• comprises three subunits: CdtA, CdtB and CdtC.
• The CdtA and CdtC subunits are responsible for mediating toxin
binding onto host cell surface receptors,
• CdtB subunit is propelled into the nucleus where it induces double-
stranded DNA cleavage, cell cycle arrest and cell death.
• C. coli, C. fetus subsp. fetus, C. fetus subsp. venerealis, C.
hyointestinalis, C. jejuni, C. lari and C. upsaliensis all harbor genes
encoding CDT

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 Cytolethal distending toxin
• CDT induces progressive cellular distension and eventual cell death,
characteristics that have been observed in CHO, HeLa, HEp-2, Vero
and chicken embryo cells inoculated with cell lysates or filtrates of C.
coli, C. fetus or C. lari
• Similarly, HeLa cells and human T cells exposed to lysate of C.
upsaliensis exhibited DNA fragmentation and failed to divide, owing to
blockage at the G2/M phase.
• 41% of 718 Campylobacter strains tested were positive for CDT
activity, including 44% of the C. lari strains (7 of 16) and 43% of the C.
fetus subsp. fetus strains (3 of 7). C. concisus is thought to secrete a
toxin similar to the CDT.

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 Cytolethal distending toxin
• CDT induces cellular distension and cell death, characteristics that
have been observed in CHO, HeLa, HEp-2, Vero and chicken embryo
cells inoculated with cell lysates or filtrates of C. coli, C. fetus or C. lari
• HeLa cells and human T cells exposed to lysate of C. upsaliensis
exhibit DNA fragmentation and blockage at the G2/M phase.
• 41% Campylobacter strains tested were positive for CDT activity,
including 44% of the C. lari strains and 43% of the C. fetus subsp.
fetus strains. C. concisus is thought to secrete a toxin similar to the
CDT.
• facilitates C. jejuni colonization, induction of intestinal inflammation
and confers resistance to bacterial clearance in different mouse
models
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 Other toxins
• A 104 kDa cytotoxin has been identified in C. rectus, which has an
amino acid composition similar to other hemolysins of Gram-negative
bacteria
• Decreases host cell viability to 57–68%.
• Cell-bound or secreted hemolysins have been reported in C. concisus
and C. coli, but not in C. lari and C. fetus subsp. fetus.
• Other cryptic Campylobacter toxins have been documented in the
literature. enterotoxins produced by C. lari strains have been shown to
induce cytotoxic and cytotonic (elongation) effects on cell lines
• enterotoxin produced by C. hyointestinalis has been reported.
• C. concisus strains isolated from healthy individuals and patients with
diarrhea harbored the zonula occludens toxin gene.
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 TSS
• The type IV secretion system (T4SS) is a machinery by which bacteria
deliver effector proteins or DNA into eukaryotic host cells or other
bacteria.
• In C. jejuni, the T4SS mediates invasion into host cells. It has been
proposed that C. fetus subsp. venerealis and C. coli also possess a
T4SS.
• C. fetus subsp. venerealis deficient in one of the T4SS-associated
genes displayed a 42% reduction in invasiveness into Caco-2 cells
and had impaired ability to induce cytotoxicity in HeLa cells as
compared with the corresponding wild-type strain.
• genomes of C. jejuni, C. coli and C. concisus contain putative genes
that might encode T6SS proteins
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 Surface Layer
• C. fetus and C. rectus are the only two members within the
Campylobacter genus identified so far to have a surface layer
(S-layer),
• a lattice that coats the outer membrane of these bacteria. S-layer
proteins of C. fetus and C. rectus have a central role in immune
evasion
• C. jejuni, C. coli and C. upsaliensis that naturally lack the S-layer are
highly susceptible to serum lysis and phagocytosis by human
polymorphonuclear cells.
• mutant deficient in an S-layer protein was rapidly engulfed by
polymorphonuclear cells within 5 min of infection.

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 Surface Layer
• Similarly, removal of S-layer proteins by a mixture of proteolytic
enzymes markedly abolished the capacity of C. fetus to resist serum
killing.
• Binding of complement C3b to the surface of wild-type C. fetus has
been shown to be markedly impaired compared with mutant C. fetus
strains lacking an S-layer protein.
• The impaired binding of stable complement C3b to the bacterial cell
surface might also affect subsequent binding by C5 and C9, which is
required for the formation of the membrane attack complex.
• the expression of S-layer proteins in C. fetus undergoes antigenic
variation in relapsing infection, which adds a layer of complexity to
S-layer-mediated immune evasion
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 Proposed mechanisms of pathogenesis

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 Diagnosis
• Campylobacter infection is seen in children and young adults and results
in acute enterocolitis characterized by severe abdominal cramp, nausea,
fever, muscle ache, headache, and acute watery to bloody diarrhea
lasting for 3–4 days.
• self-limiting; however infection can be severe and even may lead to death
in patients that are immunocompromised.
• Guillain-Barré syndrome develops several weeks after the start of the
infection characterized by generalized paralysis.
• Reiter’s syndrome also develops in patients affecting the knee joint and
the lower back.
• can cause other inflammatory conditions such as appendicitis,
endocarditis, peritonitis, meningitis, and cholecystitis.

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 Detection
• Isolation medium: Campylosel medium uses cefoperazone, vancomycin,
and amphotericin B as selective agents. The CCDA (charcoal
cefoperazone, deoxycholate agar) and CAT (cefoperazone, amphotericin
B, teichoplanin) media have been used for isolation of Campylobacter
at 37 °C under microaerophilic conditions
• PCR-based assays for detection of Campylobacter species using target
genes included flagellin (flaA), 16S rRNA, and 16S/23S intergenic spacer
region.
• Campylobacter species identification and typing has been done by using
Ribotyping, restriction fragment length polymorphism (RFLP), amplified
fragment length polymorphism (AFLP), pulsed-field gel electrophoresis
(PFGE) and randomly amplified polymorphic DNA (RAPD)-PCR
methods.
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 Treatment strategies

• Since the campylobacteriosis involves self-limiting diarrhea,

antibiotic therapy is not required,
• Maintenance of hydration and electrolyte balance is advised.
• Antibiotic therapy is however, needed for immunocompromised
patients to control bacteremia and sepsis.
• Erythromycin and newer microlides, azithromycin, and
clarithromycin are effective against C. jejuni infection.
• Increased resistance of Campylobacter to fluoroquinolone
discourages its therapeutic application.

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