CLINICAL BACTEROLOGY (LECTURE)

LESSON 10: VIBRIO, AEROMONAS, CAMPYLOBACTER AND HELICOBACTER

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2nd SEMESTER I S.Y. 2021-2022
TRANCRIBED BY: JEAN HERSHEY REYES

Vibrionaceae o Require the addition of sodium for growth

 Commonly found in a wide variety of aquatic environments, Vibrio – Antigenic Structure

including fresh water, brackish or estuarine water, and marine or
Three major subgroups of V. cholerae
salt water
 Vibrio species have been associated with the large epidemics and
pandemics
o Encompasses more than 110 validly publish species.
o 10 found in human clinical specimen.
o The risk of infection can be reduced by the avoidance of
eating raw and undercooked shellfish.
o Avoid exposure of wounds to fresh estuarine and marine
water resources.
o Cholera- 1817
o V. cholerae O1, V. cholerae O139, and V. cholerae non-O1
 V. Cholera o1 has been disease of major
public health significance of century.
o All of which share a common flagellar (H) antigen and somatic
(O) antigen
o Based on the composition of O antigen, V. cholerae O1
organisms are divided into the following serotypes:
1. Ogawa – India
2. Inaba – Philippines
3. Hikojima – Japan

 Campylobacter plays a role in GBS

 Helicobacter causes ulcer; has been linked to gastric carcinoma. Vibrio cholerae
 Most commonly isolated species:
 V. cholerae O1 – causative
o V. cholerae – cause a devastating diarrheal disease:
agent of cholera (aka Asiatic
Cholera
cholera or epidemic cholera)
o V. parahaemolyticus
o V. vulnificus
o V. alginolyticus

Vibrio – Clinical Manifestations

Factors to consider when identifying Vibrio infection:

1. Recent consumption of raw seafood (especially oysters)
2. Recent immigration or foreign travel
3. Gastroenteritis with cholera-like or rice-water stools
4. Accidental trauma incurred during contact with fresh,
estuarine, or marine water or associated products (e.g.,
shellfish, oyster or clam shells, fishhooks)
Vibrio – Microscopic Morphology
 Asporogenous
 Curved (comma) or pleomorphic gram-negative rods
 In broth – possess polar, sheathed flagella
 In solid media – peritrichous, unsheathed flagella
Vibrio – General Characteristics
 Facultative anaerobe
 Catalase negative (except for V. metschnikovii)
 Oxidase positive (except for V. metschnikovii)
 Reduce nitrate to nitrite (except for V. metschnikovii)
 Glucose Fermenter
 Vibriostatic 0/129 susceptible
o 0/129 exhibiting a zone of inhibition to 150 microgram
vibrio stat disc on Weller-Hilton/MSA
 Do not grow on MAC
 Routine media used: TCBS
 Most species exhibit a positive string test including V. cholerae
o Observed as a mucoid “stringing” reaction after
emulsification of colonies in 0.5% sodium desoxycholate
 All species are halophilic or “salt-loving” (except for V. cholerae
and V. mimicus)
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Cholera
 Spread through contaminated water
 The disease manifests in acute cases as a severe gastroenteritis
accompanied by vomiting and followed by diarrhea
 Rice watery stool (contain mucus flecks) that can rapidly lead to
dehydration and death
 Responsible: Cholera toxin, or choleragen
 Treatment and management: intravenous or oral fluids to replace
fluids lost from the severe diarrhea
o Can be shorten by antibiotics such as azithromycin or
ciprofloxacin
o Cholera is prevalent in the Bengal region of India and
Bangladesh.
o Improperly preserved and handled foods including fish,
seafood, milk, ice cream and unpreserved meat
(responsible for outbreaks)
 Liquid stools referred to as rice water stools.
 Enterotoxin (cholera toxin)
Two Types of Cholera
 Classical Type of Cholera
 El Tor Type of Cholera (more serious); it is fatal
Differentiated by:
o Voges Proskauer Test – Pos: El Tor
o Polymixin B Susceptibility Test – Susceptible: El Tor
 o Serological Test using Chicken RBC as Antigen – damsela,, and most V. furnissii, V. cincinnatiensis,
Agglutinate chicken RBCs – El Tor vulnificus strains and some V. metschnikovii
V. vulnificus strains
Other Vibrio spp.
 Enrichment procedure – alkaline peptone water with 1% NaCl
can be inoculated (at least 20 mL) and incubated for 5 to 8 hours
at 35° C
 Second most common Vibrio
 All Vibrio spp. are inositol fermenter (except V. cincinnatiensis
species implicated in gastroenteritis and some strains of V. metschnikovii)
 Most cases of gastroenteritis can be  Vibriostatic agent O/129
traced to recent consumption of raw,
improperly cooked, or Aeromonas
recontaminated seafood, particularly
Vibrio oysters Major Clinical Species:
parahaemolyticus  Self-limited disease o A. hydrophila
 Kanagawa-toxin Positive – o A. sobria
o A. caviae – most commonly isolated specie
Produces a heat-stable hemolysin
that is able to lyse human  Isolated from retail produce sources and animal meat product
erythrocytes in a special, high salt  Enteric pathogen (salmonella, Shigellae, or vibrio cholerae)
mannitol medium – Wagatsuma  Not always involve aquatic exposure such as:
Agar o An association with untreated water
 Summer Diarrhea o Consumption of raw seafood (oysters/clumps); dairy
 Pathogen in Japan products
 Causes food poisoning  24 hours incubation at 35 degree celcius
 Normal microbiota E. coli.
o Watery diarrhea
o Moderate cramps/vomiting
General Characteristics
o Fever (little)
 Symptoms: 24-48 hours  Ubiquitous
 The pathogenic vibrio usually grow  Oxidase positive- distinguish entero except Pleisomonas
as non-lactose fermenter- E.coli shigellosis (fermentation of inositol)
o Aeromonas- negative
Vibrio alginolyticus 1. Least pathogenic; most frequently
o Pleisomonas- positive
isolated
 Glucose fermenter
Vibrio vulnificus 2. Two categories of infection:
 Gram-negative rods
primary septicemia and wound
 Motile with single tuft of flagella
infections
 Widely distributed in freshwater, estuarine, and marine
environments worldwide
Laboratory Diagnosis of Vibrio spp.
Clinical Manifestations
 SBA and CAP colonies – medium to large colonies that appear
smooth, opaque, and iridescent with a greenish hue  They may cause diarrheal disease as well as other miscellaneous
o SBA - α- or β-hemolysis infections
 MAC – NLF colonies (except for V. vulnificus)  Diarrhea diarrheal infection is usually self-limiting
 Oxidase positive  A. caviae – most frequently associated with gastrointestinal
infections
 MAC and CIN agar can give false-positive oxidase reactions
 Extraintestinal infections – septicemia, meningitis and wound
 Transport media – Carry-Blair Media
o Not recommended – Buffered Glycerol Saline infections
 Most common presentation of wound infection – cellulitis
 Recommended selective medium – TCBS (thiosulfate citrate bile
salt sucrose agar)
o Differentiates sucrose-fermenting (yellow colonies) Laboratory Diagnosis
species from nonsucrose-fermenting (green colonies)
species  Colony appearance – large, round, raised, opaque colonies with
an entire edge and a smooth, often mucoid surface
o extremely strong odor is present, and pigmentation
ranges from translucent and white to buff-colored
 Major clinical species hemolytic pattern – strong β-hemolysis
 CIN colony appearance – pink-centered colonies (mannitol
fermenter)
 Inositol negative
 Oxidase positive
 Glucose fermenter with or without the production of gas
o caviae – Lactose fermenter
 String test negative
 Vibriostatic O/129 resistant
o Aeromonas- resistance
Non-sucrose Fermenter Sucrose Fermenter o Pleisomonas- susceptible

 V. mimicus, V.  V. cholerae, V.  Can grow in a media not containing salt (same with Pleisomonas)
parahaemolyticus, P. alginolyticus, V. fluvialis, V.
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 Campylobacter and Helicobacter Broth with 20% glycerol and
frozen at −70° C
Most clinically relevant species:  Commonly used selective media for
1. C. jejuni subsp. jejuni C. jejuni – Campy-BAP (blood agar
2. C. coli
plate)
3. C. fetus subsp. fetus
o Other media used:
Campylobacter Butzler medium and
Skirrow’s medium
 GBS – Guillain-Barré Syndrome (1 in 1,000 diagnosed Culture Media
campylobacter infections)  Commonly used media for H.
 C. jejuni – most common cause of bacterial gastroenteritis pylori – CAP or Brucella agar with
worldwide 5% Horse red blood cells
o Self-limited (2-6 days) (nonselective medium)
o Some is sexually transmitted disease o Selective media:
o Diarrheal disease begins with mild abdominal pain within Skirrow’s agar
2-10 days after ingestion of the organisms. o Microaerophilic
 Fever, chills, nausea and vomiting. environment

Mode of transmission:
o Direct contact with animals and handling infected pets,
such as dogs, cats, and birds
o Indirectly by the consumption of contaminated water and
dairy products and improperly cooked poultry
o Person-person transmission

 C. fetus is the causative agent of bacteremia

 Campylobacter infection plays a role in GBS, an autoimmune
disorder characterized by acute paralysis caused by damage to
the peripheral nervous system
 Does not belong with the vibrios
 Asaccharolytic
 P2 isolated from blood cultures
 Immunocompromised and elderly patients

Helicobacter

 Important specie: Helicobacter pylori – major cause of type B

gastritis or peptic ulcer
o Classified as carcinogen
o Important risk factor for gastric carcinoma – long term H.
pylori infection
o Colonizes the stomach for a long time and can cause a
low-grade inflammatory process
 Incubation temperature:
Laboratory Diagnosis o Campylobacter jejuni – 42°
C (growth of colon
 Specimen for Campylobacter microbiota is inhibited at
causing gastroenteritis (C. jejuni): this higher temperature)
 Stool o H. pylori – 37° C
 Rectal swab (less preferred) Incubation  Both Campylobacter spp. and
Helicobacter spp. require a
Specimen Collection
 Delay in processing – transport microaerophilic and capnophilic
and Transport
medium used is Cary-Blair environment
o Buffered-glycerol saline – o Campylobacter – 5% O2,
avoided 10% CO2
 Specimen of choice of C. fetus – o Helicobacter – 5% to 10%
blood with incubation at 35◦C to 37◦C O2 and 5% to 12% CO2

 Specimen for Helicobacter – gastric Microscopic  Brucella or Tryptic Soy broth

Morphology  Campylobacter spp. – curved non–
biopsy material
o Transport medium – Stuart spore-forming, gram-negative rods
medium o May appear as long
o Tissue samples may also be spirals or ‘S’ or seagull-
placed in Cysteine-brucella wing shapes

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 o Stain poorly on gram-
stained smears
 Carbol fuchsin
is recommended
as a counter
stain
o “Darting” motility
(corkscrew movement) on
hanging drop preparations
or when visualized under
phase contrast microscopy
o Single polar flagellum

 Helicobacter spp. –
morphologically similar with
Campylobacter
o Curved or U-shaped
o Motile with multiple flagella
at one pole

 C. fetus subsp. fetus – smooth,

convex, translucent colonies

 C. jejuni and other enteric

Colony Morphology Campylobacters – moist, runny
looking, and spreading
o Nonhemolytic
o Some are round and
raised and others may be
flat

 Both genus – Oxidase positive

 Urease test: Pos – H. pylori; Neg –
Definitive C. jejuni
Identification
 Hippurate hydrolysis test positive –
C. jejuni

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