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Outline Taxonomy
o I. Campylobacter spp.
A. Campylobacter jejuni
Kingdom: Bacteria
B. Other Campylobacter spp. Phylum: Proteobacteria
1. C. fetus
2. C. concisus
Class: Epsilon
3. C. fetussubspvenerealis Order:Campylobacteriales
4. C. jejunisubsdoylei
5. C. coli Family: Campylobacteriaceae
6. C. lari Genus: Campylobacter
7. C. hyointestinalis
8. C. upsaliensis A. Campylobacter jejuni
9. C. soutorum Representative of the genus
II. Helicobacter spp. Genus Campylobacter means “curved rods”
A. Helicobacter pylori
B. Other Helicobacter spp.
(campylos and baktron)
1. H. cinaedi Small, comma-shaped
III. Vibrio spp.
2. H. fennelliae
Bipolar flagella at both ends
A. Vibrio cholerae 0.2-0.9 micron in width and 0.5-5.0 micron in length
B. Halophilic Vibrios
1. V. parahaemolyticus Gram (-) bacilli
2. V. alginolyticus
3. V. vulfinicus
Motile
Resemble Vibrio species
LEGEND Pleomorphic- different shapes; if culture is old, become
Book coccoid in shape
Transers Notes Non-spore former
CAMPYLOBACTER SPP. Microaerophilic (require small amounts of oxygen,
Clinical Case # 1 usually 5 to 10 %)
A.M., a 24- year old female was brought to the ER for Thermophilic at 42° C but can also live at mesophilic
bilateral, ascending lower extremity weakness and temp.
associated dyspnea. She was conversant but began to Selective media: Butzler’s medium, Preston
breathe heavily after 2 days since hospital admission and her Campylobacter selective medium, and Blaser’s
oxygen saturation dropped from 97% to 60%. Immediately, medium (Campy-BAP) , Skirrow’s medium
she was intubated. Laboratory results revealed normal CBC Plates incubated for 48 hours
without leukocytosis and her CXR revealed normal results. A Circular and convex but can be flat and tend to swarm on
neurologist was brought into the team and assessed the case moist agar
as Guillain- Barre Syndrome (GBS). A month ago, the
Non CHO fermenters
patient complained of a 4-day history of acute diarrhea
Oxidase (+)
associated with headache, nausea, vomiting, body malaise
and undocumented fever. Previously, she was brought to the Catalase (+)
same ER but was sent home and was given probiotics. There Nitrate Reducer
was resolution of the diarrhea. What could have caused her Sodium hippurate hydrolyzer only for C. jejuni
GBS? Campylobacter jejuni: 90-95% of all human
Campylobacter infections,
Discovery
C. coli is highly homologous to C. jejuni
Tip: dahil classified sya under enterobacteriaceae, this
1886: Theodore Escherich described a non-culturable organism is oxidase, catalase positive and a nitrate reducer
spiral-shaped bacteria
How can humans be infected by C. jejuni
1909: T. Smith demonstrated sheep abortions were
associated with this infection hence named Vibrio fetus - Ingestion –fecal oral route
gastrointestinal complication would be septic abortion in Jejunum and ileum are first colonized
sheep Colon and rectum are also infected
1947: V. fetus was isolated in blood culture from a Invasive and cytopathic
pregnant patient Can cause Mesenteric adenitis- inflammation of lymph
o Developed influenza-like symptoms nodes in the mesentery; can cause appendicitis, can pass
o Stillborn fetus with necrotic and infarcted through portal vein circulation then cause sepsis
placenta Occult bacteremia- can develop into sepsis
1957: King hypothesized V. fetus can cause human
enteric disease- through zoonotic infections
o Pregnancy, gastrectomy, tooth extraction, heart
disease, DM, cirrhosis were predisposing factors
seen in association studies of 15 patients
o Butzler et al showed V. fetus in diarrheic
children’s stools
1967: Sebald and Veron separated Campylobacter from
Vibrio species and reclassified V. fetus as Campylobacter
Culture
o Stool sample or rectal swabs usually; blood
possible
0
o Campylobacter spp survive 1-2 weeks at 4 C in
Cary-Blair transport medium
o Selective media: Butzler’s selective medium,
Skirrow’s, Campylobacter selective medium, Preston
Campylobacter selective medium and Blaser’s
medium (Campy-BAP)
o Skirrow’s medium: vancomycin, polymyxin B,
trimethoprim
0 0
o Plates incubated at 42 C but C. fetus at 37 C
o O2 5%; CO2: 10%; N2:85% (microaero)
o Flat and effuse, tendency to spread on moist agar
o Nonhemolytic, colorless, moist, flat, convex D
Biochemical testing
o C. Jejuni is the only one that can hydrolize
Hippurate (Mnemonic: Camp John Hay: C. jejuni:
Virulence Factors hippurate)
Serology
Heat-labile Enterotoxin
o Complement fixation test
o Resembling cholera toxin
o ELISA
o Raises intracellular cAMP
o Causes diarrhea
Capsular polysaccharides
o Increase adherence to human enterocytes
o Provides resistance against epithelial microbial
peptides ((β-defensins, lysozymes) and lysis and
phagocytosis
o Needed in biofilm production
Flagellin A (flaA) – for motility
o Adhesion and invasion
o Overcome intestinal peristalsis
Cytolethal distending toxin (CDT)
o Best characterized toxin
o 3 subunits: cdtA and cdtC binds to host cell- trimeric
o cdtB has DNAse I-like activity- minsan nag a-
apoptosis yung cells mo
o Host cell destruction- can cause also apoptosis
Type IV (T4SS) and VI (T6SS) secretion systems
o Similar to conjugation
o Resembles the tail spike of the T4 phage
Clinical Manifestations
• Acute onset
• Crampy abdominal pain
• Profuse diarrhea- Majority of patients have this, may be
Dysenteric (bloody diarrhea)
• Headache
• Malaise Treatment
• Fever
• 5-8 days Gastroenteritis is self-limiting
• Prolonged carriage in immunodeficient patients Fluid and electrolyte management (center of treatment
• Complications: Aseptic arthritis and Guillain-Barre for general diarrhea)- to prevent hypovolemic shock
syndrome Severe gastroenteritis and sepsis treated with
• Major clinical features of campylobacter that have a erythromycin, tetracyclines, quinolones
median frequency of 84% manifest diarrhea, followed by Proper food preparation
abdominal pain then fever, myalgia, headache and lastly Pasteurization of milk
vomiting and blood in feces Prevention of contaminated water supply
Diagnosis
Clinical Microscopy
o Gram staining = gull wing shaped rods
o Dark field/phase contrast = darting or tumbling
motile spiral rods
o Stool direct smear
Cruz, De Leon Checked by: YAO Page 2 of 10
MICROBIOLOGY Campylobacter, Helicobacter, and Vibrio MODULE 3, LECTURE 3
B. Campylobacter fetus
Veterinary pathogen Differentiation between C. jejuni and C. fetus
Infective abortion in cattle and sheep
Opportunistic in systemic infections of
immunocompromised patients
Occasionally cause diarrhea
• Bacteremia, septic thrombophlebitis, arthritis, septic
abortion, meningitis
C. Campylobacter concisus
Gingivitis
Periodontal disease
Can be isolated in stool
D. Campylobacter fetus subsp. venerealis
Enzootic infertility in cattle
Not zoonotic
E. Campylobacter jejuni subsp. doylei
Does not reduce nitrate to nitrite but hydrolyzes
hippurate
Pathogenicity still unknown
Isolated from human gastric epithelium biopsy and stool
F. Campylobacter coli
Clinical infection indistinguishable to C. jejuni
3-5 % of Campylobacter diarrhea
Healthy pigs as reservoir
Hippurate hydrolysis (-)
G. Campylobacter lari
Formerly known as C. laridis
found in birds and dogs HELICOBACTER SPP.
Zoonotic Clinical Case # 2
H. Campylobacter hyointestinalis
Swine ileitis M.L., A 27- year old male from Quezon City came to
Previously nonzoonotic but reportedly found in humans your clinic complaining of an acute abdominal pain. He said
with diarrhea and homosexual men with proctitis that the pain was emanating from the epigastric area, the pain
I. Campylobacter upsalienses severity was 10/10, associated with recurrent vomiting. The
From dogs abdominal pain started 3 weeks ago when he started to be
Occasionally causes human diarrhea stressed from his PRC board exams. So, he drank a lot of
J. Campylobacter sputorum coffee to be able to study in the wee hours of night. After being
Normal flora of the respiratory tract and gingival seen by you at the clinic, he was immediately brought into the
crevices ER. CXR was done and revealed normal results. He was
Isolated in 2% of healthy people referred to GI and an EGD with gastric biopsy was done a day
Diarrhea, abscess, and sepsis after admission. The scope revealed multiple ulcerations in
the duodenum. Histopathologic biopsy specimens revealed a
spiral shaped rod bacteria near the gastric epithelium. He
said that he had recurrent abdominal pain for 5 years
already but this time, it was the worst ever he had experienced. Bacterial antigen
What could be this bacterium? o Cross-reacts with anthral gastric antigens stimulating
an autoimmune response
Discovery
1983: Warren and Marshall observed campylobacter-like Protease
bacteria in gastric mucosa of patients with gastritis and o Degrades gastric mucosa
peptic ulcer
Originally called C. pyloridis Ability to cause hypergastrinemia
Different from other campylobacters because of o Upsets gastrin-HCl homeostasis
multisheathed flagella, strong urea hydrolyis, unique fatty
acid profile
Colonized stomachs of half the world’s population
Taxonomy
Kingdom: Bacteria
Phylum: Proteobacteria
Class: Epsilonproteobacteria
Order: Campylobacterales
Family: Helicobacteraceae
Genus: Helicobacter
A. Helicobacter pylori
Gram (-) spiral-shaped
0.5 – 0.9 microns wide x 2 – 4 microns long
Motile due to tuft of sheathed unipolar flagella
Nonspore former
Pleiomorphic, old cultures are coccoid
Microaerophilic
Mesophilic (35 -37)
H. pylori Media
o Can be grown in chocolate agar or Skirrow’s
Campylobacter selective medium
o Circular, convex, translucent colonies
o 3-5 days incubation
o Biochemically, produces urease (used for rapid
Pathogenesis
diagnostic test), oxidase (+), catalase (+),
phosphatase (+), H2S (+) Gastric mucosa colonization
o Noncarbohydrate metabolizer Antrum
o Nitrate reducer (-) Present in large numbers in the mucus overlying the
mucosa at pH 7
Extends into the gastric glands
Gastric juices, bile, saliva
Bacterial protease, toxins, ammonia
H. pylori is toxic to gastric acid
Urease produces ammonia that buffers and neutralizes
the acid near the periphery of the bacterium
Chronic active or type B gastritis
Antral gastritis, duodenal and gastric ulcers, gastric
malignancies: gastric adenoCA and MALT lymphoma
Urease
o Produces ammonia that neutralized HCl
o Favors bacterial growth
Invasive Tests
Microscopy (of BIOPSY)
o Gram staining, silver staining, H and E
staining, Giemsa staining, IF
o Warthin-Starry silver stain: most
sensitive
Culture Treatment in ADULTS
o Done on nonselective medium (chocolate Triple therapy with metronidazole and either bismuth
agar) and selective medium (Skirrow) subsalicylate or bismuth subcitrate plus either amoxicillin
o Incubated from 2-7 days in a moist. or tetracycline for 14 days eradicates H pylori infection in
Microaerophilic atmosphere at 35 – 37 70–95% of patients. An acid-suppressing agent given for 4
degrees Celsius in the presence of 5 – 10 to 6 weeks enhances ulcer healing. Proton pump inhibitors
CO2; high humidity is essential (PPIs) directly inhibit H pylori and appear to be potent
o Morphology, gram staining, biochemical urease inhibitors. The preferred initial therapy is 7–10
properties days of a PPI plus amoxicillin and clarithromycin or a
Biopsy Urease Test quadruple regimen of a PPI metronidazole, tetracycline,
o Biopsy specimen is placed into a small and bismuth for 10 days
quantity of urea solution Bismuth salicylate
o An indicator of alkalinity results in ammonia Tetracycline
formation via urease Amoxicillin
o Specimen is crushed in 0.5 ml urea with Metronidazole
indicator and incubated at 37 degrees Clarithromycin o 2 weeks at least
Celsius
Proton pump inhibitor
o If bacteria are present, pH changes within a
few minutes to 2 hours due to ammonia
production
o Detects allkalinity within 2 hours
Cultural Characteristics
Aerobic
Grows in 13-40 degrees Celsius (optimum 37)
Better in alkaline medium (pH 7.4-9.6) with optimum pH
Treatment in CHILDREN at 8.2
*almost the same , varies with dosage only Nonhalophilic (does not require salt)
B. Helicobacter cinaedi Grow them in ordinary and special media
Proctitis in homosexual males Types of Media
Bacteremia in MSM with concurrent TB associated with Ordinary Media
HIV and AIDS a. Nutrient Agar
C. Helicobacter fennelliae Moist, translucent, round disks
Proctitis in homosexual males 1-2 mm, bluish tinge
D. Helicobacter heilmanii Distinctive odor
Spirochetal in morphology b. Mac Conkey Agar
Strongly urease (+) Colonies are smaller
Occurs in 1% in the gastric mucosa Colorless but become reddish due to late
VIBRIO SPP. lactose fermentation
Clinical Case # 3 c. Blood Agar
C.17-year old male who was severely emaciated Colonies are surrounded by green zone due
and had diarrhea for only a day but had 25 bouts of to hemodigestion
diarrhea. The volume of every stool was approximately half a d. Gelatin Slab
pail. The stool looked like washings of rice. The senior doctor White line of growth along the track of the
with you in the medical mission ordered for an IV drip and an inoculating wire
IV antibiotic. A week after, the patient was already well and
Liquefaction of gelatin begins at the top which
was discharged uneventfully. What could have caused this
spreads in a funnel-shaped (infundibuliform)
patient’s diarrhea?
or turnip shaped (napiform) manner in 3 days
Introduction at 22 degrees Celsius
12 out of the 35 Vibrio spp. are pathologic e. Peptone Water
Halophilic and nonhalophilic Incubated at 37 degrees Celsius
Grow with added salt (HALOPHILIC) Fine surface pellicle which breaks into
Vibrio cholerae: representative of the genus membranous pieces after shaking
– causes cholera Turbidity and a powdery deposit
– 1883: Isolated in Egypt by Robert Koch
V. parahaemolyticus, V. vulnificus, V. minicus, V. Special Media
alginolyticus f. Holding or Transport Media
A. Vibrio cholera Venkatraman-Ramakrishnan (VR)
Morphology o 20 g crude sea salt + 5 g peptone in 1 L
Gram (-), short, curved, cylindrical rods distilled water with pH 8.6-8.8
1.5 micron x 0.2-0.4 microns in size o Screw capped im 10-15 ml
o 1-30 ml stool added
Rounded or slightly pointed ends
o Vibrio does not multiply but remain
V. comma
viable for several weeks
S-shaped or spiral forms seen when 2 or more cells lie o Prevents overgrowth
end to end
Upon serial transfers: become straight
Pleiomorphic in old cultures
Biotypes Pathogenesis
2 biotypes: classical and El Tor Fecal-oral route