BACT211: CLINICAL BACTERIOLOGY
TOPIC: VIBRIO, AEROMONAS, AND CAMPYLOBACTER SPECIES
2ND SEMESTER | S.Y 2023-2024
LECTURER: Ma’am Christy Gonzales
TOPIC
SUBTOPIC
SUB SUBTOPIC
VIBRIO
• Family Vibrionacceae
• Asporongenous, gram-negative rods that measure
approximately 0.5 to 0.8 m in diameter by 1.4 to 2.6
m, facultative anaerobic, curved or comma-shaped
rods
• Motile, catalase, and oxidase positive
• Gram negative bacilli, curved or comma-shaped rods,
facultative anaerobes, and motile
- Could have monotrichous flagella or peritrichous
• Able to reduce nitrate to nitrate except V.
metschnikovii [not part of the normal flora; can be
found in the environment, mostly water]
• Found in marine and surface waters
• Can grow within a broad temperature range (14-40°C)
• Halophilic except for V. cholerae and V. mimicus
• Susceptible to the vibriostatic compound O/129 (2,4
diamino-6,7-diisopropylpteridine)
• Positive string test – mucoid “stringing” reaction
VIBRIO CHOLERAE
• Cause of cholera
- “rice water stool”
- Diarrheal fluid loss: >1 L/h
• Cholera toxin, or choleragen [powerful enterotoxin;
virulence factor] – heat-labile enterotoxin
- Once ingested, the bacteria will colonize our small
intestine where they’ll multiply and produces
choleragen.
• Curved or Comma-shaped, curved rod 2-4 m long
• Motile-polar flagellum [monotrichous, peritrichous,
lophotrichous]
• Grows in brackish and marine waters
• Not part of our normal flora.
- Similar to N. gonorrhoeae, whenever this is present
in our body, this will cause infection
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• The basis of our classification or categorization of Vibrio
cholerae is the presence or differences of the O antigen
- V. cholerae O1, V. cholerae 0139, V. cholerae non-
O1 share a common flagellar (H) antigen and
somatic (O) antigen
o Iba iba ang kanilang O antigen
ANTIGENIC STRUCTURE AND BIOLOGICAL
CLASSIFICATION
• O Lipopolysaccharides – O1 and O139 cause
epidemic and pandemic cholera
A. V. cholerae O1
- Ogawa (A,B)
- Inaba (A,C)
- Hikojima (A,B,C)
B. V. cholerae O139
C. V. cholerae non-O1 – resemble V. cholerae but
fail to agglutinate [does not agree] in O1 antisera
• Two biotypes of epidemic V. cholerae
- Classic – does not produce hemolysis [chicken
RBC]; negative in VPT, and susceptible in
polymyxin B
- El tor – produces a hemolysin, gives positive
result on the Voges-Proskauer test, and is
resistant to polymyxin B
VIRULENCE FACTORS
• Cholera toxin (CT)
• zonula occludens (Zot) toxin (enterotoxin)
- disrupts the tight junction of our intestinal cells,
effectively decreasing tissue resistance
• accessory cholera enterotoxin (Ace) toxin
• O1 and O139 somatic antigens
• hemolysin/cytotoxins
• Motility and chemotaxis
- Being motile is a virulence factor because it has
the capability to be distributed in our body,
depending on where the signal of the food or
other organisms; pwede kumalat
• Mucinase production
- Mucinase is important in penetration in mucous
layers; small intestines can be colonized and
infected by V. cholorae
• toxin coregulated pili (TCP) pili
Transcribed by: JAMARA
 - provide means for the bacteria to attach to the
mucosal cells
VIBRIO PARAHAEMOLYTICUS
• Primary cause of so-called summer diarrhea in Japan
- Usually acquired during the summer season
• Haemophilic bacterium
• Causes acute gastroenteritis after ingestion of
contaminated seafood such as raw fish or shellfish
• Incubation period: 12-24 hours
• V. parahaemolyticus serotype O3:K6 = implicated in
numerous food-borne outbreaks in various parts of the
world
• Halophilic requirement: 1% to 8% NaCl
• Facultative anaerobe, Gram-negative rod
• Does not grow well on some of the routine differential
media used to grow salmonellae and shigellae, but it
does grow well on blood agar
• Grows well on TCBS agar [Thiosulfate Citrate Bile Salt
Sucrose] – green colonies (does not ferment sucrose)
- pH indicator: Bromthymol Blue
- Acid: yellow colonies [utilized sucrose]
CLINICAL MANIFESTATIONS
• Gastrointestinal disease – self-limited [gumagaling
ang patient without any other medication; manifest
signs and symptoms 24-48 hrs. after ingestion of
contaminated seafood]
- Kanagawa phenomenon – Heat stable hemolysin
produced by V. parahaemolyticus that is able to
lyse human erythrocytes [RBC] in a special, high-
salt mannitol medium (Wagatsuma agar)
VIBRIO VULNIFICUS
• Vibrio vulnificus can utilize/ferment lactose
- Most vibrios only ferment sucrose
• Vibrio doesn’t only cause gastrointestinal infection, it
can also cause septicemia and wound infections.
• Secondary to Vibrio cholerae, this is the second most
common organism that causes septicemia.
• Infections include:
- primary septicemia and
- wound infections (aquatic wound that often
presents as a cellulitis)
• TOC: fluroquinolones, third-generation cephalosporins
(e.g. ceftriaxone), and doxycycline
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VIBRIO ALGINOLYTICUS
• Vibrio alginolyticus, if associated with infections, they
are associated in wound infections and eye infections
• Least pathogenic for humans
• Strict halophile: requiring at least 1% NaCl
• able to tolerate up to 10% NaCl.
• Occupational hazard: people in constant contact with
seawater, such as fishermen or sailors
LABORATORY DIAGNOSIS
SPECIMEN COLLECTION
• Stool: inoculated within 2-4 hours of collection onto
appropriate agar media [As soon as possible]
• For delayed processing: mixed in a Cary-Blair
transport medium and refrigerated
- If suspecting the presence of vibrio in stool, we
will be using APW [Alkaline Peptone Water].
- Stools are non-sterile [maraming normal flora].
Those normal flora are almost all gram negative
bacilli.
- We need APW as an enrichment medium [have a
characteristic that can support the growth of
vibrios, while not supporting the growth of other
gram negative bacilli; mataas ang pH; mataas
ang sodium chloride content]
• Buffered glycerol saline – not recommended as a
transport or holding medium
- Glycerol is toxic for vibrios
DIRECT MICROSCOPIC EXAMINATION
• Gram negative bacilli
• Dark-field or phase-contrast microscopy
• “shooting star” motility: V. cholerae O1
- To easily identify, we will be using a dark-field or
phase-contrast microscopy to see the motility
[hindi makikita nag motility sa light microscope]
CULTURE
• Nutrient Agar or SBA (0.5% NaCl)
• MacConkey Agar – non-fermenters
- Lactose content [non-lactose fermenters except
V. vulnificus] [V. vulnificus will be pink; others:
colorless]
• Thiosulfate Citrate Bile Salt Sucrose (TCBS) agar
- Selective and differential medium
- As a differential, it has sucrose. [pH indicator:
Bromthymol Blue; yellow colonies when the
sucrose is fermented]
Transcribed by: JAMARA
 - It is selected because it has sodium thiosulfate,
citrate, and oxgal [bile salt; can inhibit gram
positive cocci, gram negative bacilli that are not
commonly seen in stool], high pH [can be
tolerated by vibrio]
• Alkaline peptone broth or water, containing 1% NaCl
with a pH of 8.5
- Enrichment medium for vibrios
• Taurocholate peptone broth (pH, 8.0-9.0)
• CHROMagar Vibro – V. cholerae, V. parahemolyticus,
and V. vulnificus
• V. cholerae
• V. alinolyticus
SUCROSE-FERMENTING • V. fluvialis
(YELLOW COLONIES) • V. furnissil
SPECIES • V. cincinnatiensis
• V. metschnikovii
• Some V. vulnificus
• V. mimicus
• V. parahaemolyticus
• P. damsela
(Photobacterium)
NONSUCROSE- • Most V. vulnificus
FERMENTING (GREEN) strains AEROMONAS
SPECIES - Not all Vibrio spp.
grow on TCBS, • Enterobacteriaceae family
especially - Enteric bacilli
Grimontia hollisae - Enteric gastrointestinal bacteria
(formerly Vibrio) • Vibrio spp., Aeromonas spp., Pleisiomonas spp.
COLONY APPEARANCE - Non-enteric gastrointestinal bacterial
• Medium to large, smooth, opaque, iridescent with a - Not part of the normal flora; commonly found in the
greenish hue; environment [water bodies]
• V. cholerae, V. fluviasis, and V. mimicus can be beta- - Pleisiomonas spp. is part of the Enterobacteriaceae
hemolytic NLF except V. vulnificus, which may be LF family and is oxidase positive
- In suspecting gram negative bacilli, lagi nating • Oxidase-positive, glucose fermenting, gram-negative
ginagamit ‘yung McConkey. Sa Gram positive rods
[BAP]
• May resemble morphologically members of the family
IDENTIFICATION
of Enterobacteriaceae
• Oxidase test
• Grows well on blood agar – B-hemolytic
• Nutrient broth with 6% salt – to differentiate V.
cholerae, V. mimicus, and Aeromonas spp. • A. caviae – cause of gastroenteritis, ranging from watery
• String test – to differentiate Vibrio spp. [susceptible] diarrhea (most common) to dysentery-like illness
from Aeromonas spp. - Has blood, mucus, pus
• [resistant] • A. hydrophilia – cause of wound infections; traumatic
• Vibrio static test 0/129 (2,4-diamino-6,7- soft tissue injuries
disopropylpteridine) – to differentiate vibrios - Described in patients with hematologic
(susceptible) from other oxidase positive glucose malignancies [leukemia, anemia] and/or liver
fermenters (resistant) and to differentiate V. cholerae disease
O1 and non-O1 (susceptible) from other Vibrio spp. • TOC: third-generation cephalosporins, aztreomam, and
(resistant) carbapenems, as well as aminoglycosides
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 INTESTINAL INFECTIONS LABORATORY DIAGNOSIS
• Acute secretory diarrhea – often accompanied by
vomiting CULTURE MEDIA
• Acute dysenteric form of diarrhea – similar to • SBA – strong B-hemolysis
shigellosis, with blood and mucus - A. hydrophila, A. veronii biovar sobria, and A.
• Chronic diarrhea – usually lasting more than 10 days jandaei
• Cholera-like disease – including rice water stools • MAC – Ferments lactose
• Nebulous syndrome – commonly referred to as • Modified cefsulodin Irgasin-novobiocin (CIN) II plate
traveler’s diarrhea (similar to enterotoxigenic E. coli). • CIN Medium
- Kadalasan sa immunocompromised patients - Difference: CIN II: has 4 ug of cefsulodin; CIN
nangyayari [pediatric patients, nagrereceive ng medium: 15 ug of cefsulodin
antibiotic, etc.] - Though mababa ang cefsulodin ng modfied
EXTRAINTESTINAL INFECTIONS cefsulodin Irgasin-nobociocin, mas mataas ang
yield ng aeromonas dito dahil may ampicillin sa
• Septicemia
CIN II [walang ampicillin sa CIN medium]
• Wound infections
• Pink-centered colonies from the fermentation of
- septicemia, meningitis, and wound infections,
mannitol, with an uneven, clear apron resembling
osteomyelitis, pelvic abscesses, otitis, cystitis,
Yersinia enterocolotica
endocarditis, peritonitis, cholecystitis, keratitis
PRESUMPTIVE IDENTIFICATION
associated with contact lens wear
• Oxidase Test: (Aeromonas = positive)
• Spot indole test [positive]
A. CAVIAE • Ability to grow in the presence of NaCl [hindi ganun
kataas]
• gastrointestinal infections, especially in neonate and • String Test [negative]
pediatric populations
• it has been associated with inflammatory bowel disease.
ADDITIONAL INFORMATION
A. HYDROPHILA
VIBRIO AEROMONAS PLEISIOMONAS
• most common species that can cause human Curved Gram
gastroenteritis - - -
bacilli
Motile + + +
A. VERONII (BIOVARS SOBRIA AND VERONII) Oxidase + + +
• hemolytic-uremic syndrome or kidney disease that Non-enteric
+ + +
might require kidney transplantation bacilli
• Correct writing: Aeromonas veronii biovar sobria; Gastro-
+ + +
enteritis
Aeromonas veronii biovar veronii
O129 S R S
A. VERONII BIOVAR SOBRIA Inositol
- - +
fermentation
• linked to cholera-like disease characterized by Indole - + -
abdominal pain, fever, and nausea. Mannitol + - -
6.5% NaCl + - -
TCBS + - -
Gelatin
+ + -
liquefaction
Glucose + + +

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 CAMPYLOBACTER LABORATORY DIAGNOSIS
• Strict anaerobes, microaerophilic environments SPECIMEN COLLECTION
- Requiring reduce amount of oxygen • Stool samples
• Curved, comma-, or S-shaped, Gram-negative, non- • Rectal swabs – less preferred
spore-forming rods; “sea gull wing” shapes • Transport Medium: Cary-Blair
• Primary zoonotic organisms
- Known to cause domestic abortion in animals • Helicobacter pylori: can be recovered from gastric
• Patients suffering from Guillain-Barre Syndrome (GBS) biopsy materials
• cysteine-Brucella broth with 20% glycerol and Frozen
often test positive for Campylobacter antibodies
at −70° C
• most common cause of bacterial - may laman na 10% Sheep’s RBC, vancomycin,
gastroenteritis trimethoprim, polymyxin B, amphotericin B,
CAMPYLOBACTER cephalothin
• Cramps and bloody diarrhea
JEJUNI • Stuart Medium
- Very common cause of
diarrhea in humans CULTURE MEDIA
• has been isolated most • Campylobacter blood agar or Campy blood agar
CAMPYLOBACTER • Butzler medium
frequently from blood cultures
FETUS SUBSP.
• rarely associated with • Skirrow’s Medium
FETUS
gastrointestinal illness. • Campy-CVA (cefoperazone-vancomycin-amphotericin
B) medium
- Reported that it can provide better suppression
HELICOBACTER of fecal normal flora
- 37 degree Celsius for better recovery of
• Gram negative bacilli, microaerophiles, not causing
campylobacter
gastroenteritis
• Charcoal cefoperazone deoxycholate agar (CCDA)
• strongly associated with gastric, peptic, and duodenal • Charcoal-based selective media (CSM)
ulcers as well as with gastrointestinal carcinoma.
• major cause of type B gastritis

• have been associated with

human gastroenteritis,
H. CINAEDI AND H.
generally in
FENNELLIAE
immunocompromised
patients.
H. CANADENSIS, H.
CANIS, H. PULLORUM, • gastroenteritis
AND H. • Most common: H. pylori
WINGHAMENSIS

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 INCUBATION
• 42°C
• 37°C – C. fetus subsp. Fetus

• For Campylobacter spp.

- 5% O2
- 10% CO2
- 85% N2

• For Helicobacter spp.

- 5% to 10% O2
- 5% to 12% CO2
COLONY MORPHOLOGY
• Moist runny looking, and spreading, usually
nonhemolytic, some are round and raised and others
may be flat
• C. fetus subsp. fetus – produces smooth, convex,
translucent colonies
• C. mucosalis and C. hyointestinalis – can produce a
dirty yellow pigment
MICROSCOPIC MORPHOLOGY
• Curved, non-spore-forming, gram-negative rods
• Long spirals or S- or seagull wing shapes –
campylobacter
• “darting” motility – campylobacter; phase-contrast
microscope; hanging drop preparation
DEFINITIVE IDENTIFICATION
• Positive oxidase
• Positive hippurate hydrolysis – C. jejuni

• H. pylori
- Rapid urease production/activity
- Christensen’s urea medium [slant or broth] and
incubated at 37°C for 2 hours [once there is
urease activity, nagiging salmon pink]
• Non-invasive indirect test
- Urea Breath Test
- patient will be given 13C labelled or 14C labeled
urea (ingested orally). Then, the urea will be
degraded by the urease in the gastrointestinal
tract (activity H. pylori). Sa stomach, mar-release
ang 13 Carbon Dioxide or 14 Carbon Dioxide and
maddetect kapag nagexhale ang patient sa
scintillation counter

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