CAMPYLOBACTER

 Include Helicobacter and Wolinella

 Asaccharolytic
 positive oxidase
Based on ribosomal RNA (rRNA)
sequence studies, Wolinella recta and
Wolinella curva were transferred to the
genus Campylobacter as C. rectus and C.
curvus.
 strict anaerobes, they have been
grown in a microaerophilic
environment (require oxygen, but at a
concentration less than that of room
air; 5% is normally optimal.) CLINICAL MANIFESTATIONS

EPIDEMIOLOGY
 known to cause abortion in domestic
animals
 the most common cause of bacterial
gastroenteritis worldwide (C. jejuni)
 ranking fourth in most common
causes of foodborne gastrointestinal
illness (C. coli and C. lari)
MOT: direct contact with animals and
handling infected pets
Indirectly by the consumption of
contaminated water and dairy products
and improperly cooked poultry
also sexually transmitted.
Campylobacter fetus subsp. fetus has
been isolated most frequently from blood
cultures and is rarely associated with
gastrointestinal illness. They occur in
immunocompromised and elderly
patients.
Helicobacter pylori Common cause of
duodenal ulcers and type B gastritis;
possibly a risk factor in gastric carcinoma
 fresh groundwater is the likely source
of many infections.
Campylobacter jejuni present a diarrheal
disease:
 mild abdominal pain within 2 to 10
days after ingestion of the
organisms.
 Cramps and bloody diarrhea often
follow
 Untreated patients can remain
carriers for several months.
Other enteric Campylobacter infections
(C. coli and C. lari) have similar clinical
manifestations.
Campylobacter jejuni also plays a role in
GBS. An autoimmune disorder
characterized by acute paralysis caused
by damage to the peripheral nervous
system.
 Many patients with GBS test
positive for antibodies to
Campylobacter
 Antibodies produced during a
Campylobacter infection bind to
gangliosides found on peripheral
nerves
Helicobacter pylori primarily linked to
gastric infections and classified as a
carcinogen.
 It colonizes the stomach for a long
time and can cause a low-grade
inflammatory process, producing a
chronic superficial gastritis.
also recognized as a major cause of type
B gastritis, a chronic condition formerly
associated primarily with stress and
chemical irritants.
an important risk factor for gastric
carcinoma
H. cinaedi (isolated from the blood of
patients with bacteremia and patients
with HIV infection) and H. fennelliae
have been associated with human
gastroenteritis, generally in
immunocompromised patients.
H. canadensis, H. canis, H. pullorum,
and H. winghamensis other causes of
gastroenteritis.
CLINICAL BACTERIOLOGY 1
(LECTURE): CAMPYLOBACTER
SPECIES
Laboratory Diagnosis
Specimen Collection and Transport
 C. fetus subsp. fetus can be recovered
in several routine blood culture
media.
 Campylobacter spp. that causes enteric
illness are isolated from stool samples
and rectal swabs, the less preferred
specimen.
 If a delay in processing the stool
specimen is anticipated, it can be
placed in a transport medium such as
Cary-Blair to maintain the viability
of the organisms.
 A common stool transport medium,
buffered glycerol saline, is toxic to
enteric campylobacters and should
therefore be avoided
H. pylori
 An enriched selective agar, Campy-
BAP (blood agar plate), is a
commonly used medium to isolate C.
jejuni and other enteric
campylobacters.
o It contains Brucella agar
base, 10% sheep red blood
cells, and a combination of
antimicrobials—vancomycin,
trimethoprim, polymyxin B,
amphotericin B, and
cephalothin
 Other selective media that have been
successful in recovering
Campylobacter spp. are Butzler
medium and Skirrow’s medium.
 Medium V inhibits normal colon
microbiota better than the original
formulation.
o It is a modification of the
original Butzler medium.

 Can be recovered from gastric biopsy

materials.
 Samples must be transported quickly
to the laboratory.
 Stuart medium can be used to
maintain the viability of the
organisms if a delay in processing is
anticipated.
 Tissues samples may also be placed
in cysteine-Brucella broth with 20%
glycerol and frozen at -70°C.  Incubation at 37°C allows the
recovery of Campylobacter spp. that
are inhibited at 42°C.
 C. fetus subsp. fetus, C. rectus, and
C. curvus can be isolated using
Culture Media routine culture media.

 Charcoal based, blood-free media,
such as charcoal cefoperazone
deoxycholate agar, are also available.
Image A: No sample has been
applied onto the plate
Image B: Campylobacter jejuni sbsp.
jejuni has been cultivated on the
plate. The colonies appear as
greyish to white.
 To recover H. pylori, a combination
of a nonselective medium, such as
CHOC agar or Brucella agar with
5% horse red blood cells, and a
selective medium, such as Skirrow’s
agar, may be used.
GasPak EZ Gas Generating Container
 It is important that the inoculated
medium be fresh and moist and that
the culture be incubated in a
microaerophilic environment, with
increased humidity.
Incubation
 There is a double purpose for
incubating stool cultures at 42°C to
recover C. jejuni. First, C. jejuni and
other enteric campylobacters grow
optimally at 42°C. Second, growth of
colon microbiota is inhibited at this
higher temperature.
 C. fetus subsp. fetus, on the other
hand, is a rare stool isolate, and
growth is suppressed at 42°C;
therefore, to isolate this organism,
media should be incubated at 37°C.
 Enteric Campylobacter and
Helicobacter spp. require a
microaerophilic and capnophilic
environment.
 The ideal atmospheric environment
for these organisms is a gas mixture
of:
 Except for C. rectus and C. curvus, a
strict anaerobic environment does
not support the growth of most
Campylobacter spp.
System (BD Diagnostic Systems)
 Several methods can be used to
obtain the required environment for
campylobacters. With the GasPak
EZ Gas Generating Container
System (BD Diagnostic Systems,
Sparks, MD), specimen plates along
with a sachet are placed into a clear
plastic incubation container, sealed,
and incubated at the appropriate
temperature.

Presumptive Idenfitfication
Microscopic Morphology.
Colony Morphology
 Campylobacter spp. are curved, non–
spore-forming, gram-negative rods
that measure approximately 0.2 to
0.9 µm × 0.5 to 5.0 µm
 Enteric campylobacters may appear
as long spirals or ‘S’ or seagull-wing
shapes. These organisms may appear
as coccobacilli in smears prepared
from older cultures. On Gram-
stained smears, these organisms stain
poorly.
 For better visualization,
carbolfuchsin is recommended as a
counterstain; if safranin is used,
counterstaining should be extended
to 2 to 3 minutes.
 They exhibit a characteristic
“darting” motility on hanging drop
preparations or when visualized
under phase contrast microscopy.
o It is also called shooting star
motility because of its rapid
motion that often no change
is observed in the position of
the bacterium.
 To observe the typical motility,
organisms should be suspended in
Brucella or tryptic soy broth.
Distilled water and saline seem to
inhibit motility.
 Arcobacter spp. have a microscopic
morphology similar to that of
Campylobacter spp. H. pylori also
appears similar to campylobacters,
but one ultrastructural study has
shown that Helicobacter has multiple
flagella at one pole, unlike the single
polar flagellum of campylobacters
 The typical colony morphology of C.
jejuni and other enteric
campylobacters is moist, runny
looking, and spreading.
 Colonies are usually nonhemolytic;
some are round and raised and others
may be flat.
 C. fetus subsp. fetus produces
smooth, convex, translucent colonies
 A tan or slightly pink coloration is
observed in some enteric
campylobacter colonies.
 Other Campylobacter species
produce colonies similar to those of
C. jejuni.
o Although most do not
produce pigment, C.
mucosalis and C.
hyointestinalis can produce a
dirty yellow pigment
Definitive Identification
 Isolates from stool specimens and
rectal swabs can be presumptively
identified as Campylobacter spp. by
a positive-oxidase, the characteristic
Gram-stained microscopic
morphology, and the characteristic
motility.
 The microscopic morphology is
important because it differentiates
Campylobacter from other bacteria,
such as Aeromonas and
Pseudomonas, which are oxidase-
positive and can grow at 42°C in a
microaerophilic environment.
 Here are the lists of biochemical tests
most useful for definitively
 identifying the most commonly
encountered Campylobacter,
Helicobacter, and Arcobacter
species.
 Helicobacter infections usually are
identified by nonculture methods. H.
pylori can be presumptively
identified in a gastric biopsy
specimen by testing for the presence
of a rapid urease reaction
Figure 1 Urease reactions. From Left to right,
Negative, Weak positive, Positive
 The collected tissue sample is placed
onto Christensen’s urea medium and
incubated at 37°C for 2 hours.
 A color change suggests the
presence of H. pylori.
Immunologic Assays
 Latex agglutination tests are
available for the rapid identification
of colonies of enteric campylobacters
on primary isolation media.
 Two commercial kits are available in
the United States for culture
identification: INDX-Campy and
Dry Spot Campylobacter test kit.
o These kits can detect the
presence of C. jejuni, C. coli,
C. upsaliensis, C. lari, and
sometimes C. fetus subsp.
fetus.
o However, neither system
differentiates among the
Campylobacter spp.
Commercial kits.
 Specific antibodies in serum can be
detected by enzymelinked
immunosorbent assay or indirect
immunofluorescent assay methods.
o These methods have been
reported to be reasonably
sensitive and specific
indicators of Campylobacter
and H. pylori infections.
 Serologic testing is useful for
epidemiologic studies for
Campylobacter but is not
recommended for routine diagnosis.
Serologic testing is an important
screening method for the diagnosis
of H. pylori infection.
 Antimicrobial Susceptibility

 Antimicrobial susceptibility testing

for Campylobacter spp. is not
routinely performed in the clinical
microbiology laboratory and is not
standardized
 The drugs of choice for treating
intestinal campylobacteriosis are
azithromycin and erythromycin,
although most patients recover
without antimicrobial intervention.
 Ciprofloxacin and other quinolones
can also be used.
 Gentamicin is used to treat systemic
infections. Tetracycline,
erythromycin, and chloramphenicol
can be substituted for gentamicin.
 The standard therapy for treating H.
pylori infections consists of a
macrolide, amoxicillin, and proton
pump inhibitor.
 An alternative regimen consisting of
metronidazole, tetracycline, and
bismuth salt can be used.
 Treatment for H. pylori infections
should be administered for 7 to 14
days to eradicate the infection.