Vibrio spp., Aeromonas spp., Plesiomonas spp., Campylobacter spp., Helicobacter spp., Flashcards
In terms of the morphology, the common denominator between Vibrio spp., Aeromonas spp., Plesiomonas spp., Campylobacter spp., and Helicobacter spp. is that they are all ___________.
Gram negative (-) CURVED bacilli
Vibrio spp., Aeromonas spp., Plesiomonas spp., Campylobacter spp. are all members of the __________.
Non-Enteric Gastrointestinal Pathogens
Non-Enteric Gastrointestinal Pathogens are ________.
not part of the human normal flora
Enteric Gastrointestinal Pathogens
Enterobacteriaceae
Enteric Gastrointestinal Pathogens are _______.
part of the human normal flora
Vibrio spp. came from the Family ________.
Vibrionaceae
TRUE OR FALSE.
Vibrio spp. are asporogenous.
True
Measurement in diameter of Vibrio spp.
0.5 - 0.8 um by 1.4 - 2.6 um
Aerotolerance of Vibrio spp.
Facultative Anaerobes
Specific morphology of Vibrio spp.
curved or comma-shaped rods
Are Vibrio spp. motile?
Yes
Describe the flagella of Vibrio spp.
Monotrichous or Peritrichous
Vibrio spp.
Oxidase test
Positive (+)
Vibrio spp.
Catalase Test
Positive (+)
Other gram negative (-) curved bacilli motile organisms
- Aeromonas spp.
- Plesiomonas spp.
Vibrio spp.
Reduction of Nitrate to Nitrite
Positive (+)
EXCEPT: V. metschnikovii
Which Vibrio sp. cannot reduce nitrate to nitrite?
Vibrio metschnikovii
Vibrio spp. are found in ________.
marine and surface waters
Vibrio spp.
Optimum Growth Temperature
14 - 40 C
Are Vibrio spp. halophilic?
Yes
EXCEPT: V. cholerae and V. mimicus
Vibrio spp. are susceptible to ________.
Vibriostatic Compound O/129 (2,4-diamino-6, 7-diisopropylpteridine)
Vibrio spp. are ________ to vibriostatic compound O/129.
susceptible
Vibrio spp.
String Test Result
Positive (+)
It is the most common Vibrio sp.
Vibrio cholerae
Vibrio cholerae is the causative agent of _______.
Cholera
Cholera is characterized by ________.
rice water stool
Cholera can cause diarrheal fluid loss which measures _______.
> 1 L/hour
Vibrio cholerae measures _________.
2 - 4 um long
Describe the flagellum of Vibrio cholerae
Polar Flagella
If V. cholerae is cultured in broth mediums, the flagella is _____.
polar sheathed
If V. cholerae is cultured in solid mediums, the flagella is _____.
unsheathed (peritrichous)
Vibrio cholerae
Virulence Factor/s
Cholera toxin (choleragen)
A powerful and heat-labile enterotoxin.
Choleragen
Choleragen is a powerful and ______ enterotoxin.
heat-labile
How is choleragen virulent?
Once it is ingested, the bacteria will colonize the small intestine and produce choleragen.
Which Vibrio spp. are not halophilic?
- V. cholerae
- V. mimicus
The presence or differences of this antigen is the basis of classification for Vibrio cholerae.
O lipopolysaccharide (O antigen)
It causes the epidemic and pandemic cholera.
O1 and O139
Vibrio cholerae
Three kinds of O antigen
- V. cholerae O1
- V. cholerae O139
- V. cholerae non-O1
V. cholerae O1 is further subclassified into _______.
- Ogawa
- Inaba
- Hikojima
V. cholerae O1
Ogawa O-antigen
A, B
V. cholerae O1
Inaba O-antigen
A, C
V. cholerae O1
Hikojima O-antigen
A, B, C
It resembles V. cholerae but fail to agglutinate in O1 antisera.
V. cholerae non-O1
All types of antigen share a common _______ and ________.
Flagellar (H) Antigen and Somatic (O) Antigen
Two biotypes of epidemic V. cholerae
- Classic
- El Tor
Two Biotypes of Epidemic V.cholerae
It is able to produce a hemolysin.
El Tor
Two Biotypes of Epidemic V.cholerae
It is unable to produce a hemolysin.
Classic
Two Biotypes of Epidemic V.cholerae
It is (+) for Voges-Proskauer Test.
El Tor
Two Biotypes of Epidemic V.cholerae
It is (-) for Voges-Proskauer Test
Classic
Two Biotypes of Epidemic V.cholerae
It is resistant to polymixin B.
El Tor
Two Biotypes of Epidemic V.cholerae
It is susceptible to polymixin B.
Classic
Two Biotypes of Epidemic V.cholerae
It uses chicken RBC
Classic
Enumerate the virulence factors of Vibrio cholerae
- Cholera toxin (choleragen)
- Zonula occludens toxin (ZOT)
- Accessory cholera enterotoxin (ACE)
- O1 and O139 somatic antigen
- Hemolysin/cytotoxin
- Motility
- Chemotaxis
- Mucinase
- Toxin coregulated pili (TCP)
Virulence Factors
It disrupts the tight junctions in the intestinal cells, decreasing the tissue resistance.
Zonula Occludens Toxin (ZOT)
Virulence Factors
It allows the bacteria to be distributed in the body.
Motility
Virulence Factors
It allows for the penetration to mucus layers.
Mucinase
Virulence Factors
It attaches the bacteria to the mucosal cell.
Toxin Coregulated Pili (TCP)
It is the primary cause of summer diarrhea in Japan.
Vibrio parahaemolyticus
V. parahaemolyticus is the primary cause of _______ in Japan.
summer diarrhea
V. parahaemolyticus is a halophilic bacterium that requires _______.
1 - 8% NaCl
It causes acute gastroenteritis after ingestion of contaminated seafood.
Vibrio parahaemolyticus
V. parahaemolyticus causes ________ after ingestion of contaminated seafood.
Acute gastroenteritis
Acute gastroenteritis can be acquired from ______.
Eating contaminated seafood
Vibrio parahaemolyticus
Incubation Period
12 - 24 hours
A serotype of V. parahaemolyticus which is a pandemic type, and implicated in food-borne outbreaks in the world.
O3:K6
Vibrio parahaemolyticus
Aerotolerance
Facultative Anaerobes
TRUE OR FALSE.
V. parahaemolyticus grows well on blood agar.
True
TRUE OR FALSE.
V. parahaemolyticus does not grow well on TCBS agar.
False
Vibrio parahaemolyticus
TCBS Result
Green Colonies
They do not ferment sucrose.
pH indicator for TCBS agar
Bromthymol blue
(+) result for TCBS agar
Acid (+): Yellow colonies
Sucrose Fermenters
Vibrio parahaemolyticus
Clinical Manifestations
Gastrointestinal Disease (self-limited)
A special, high salt mannitol medium.
Wagatsuma Agar
A characteristic possessed by V. parahaemolyticus which enables it to lyse RBCs in a special, high salt mannitol medium.
Kanagawa Phenomenon
Explain the Kanagawa Phenomenon
V. parahaemolyticus is a heat-stable hemolysin that is able to lyse RBCs in a special, high salt, mannitol medium called Wagatsuma Agar.
Vibrio parahaemolyticus
The signs and symptoms for gastrointestinal disease usually appears after ______.
24-48 hours of ingestion of contaminated seafood
It is the second most common cause of septicemia.
Vibrio vulnificus
Infections caused by V. vulnificus
- Primary Septicemia
- Wound infections
Vibrio vulnificus
Treatment of Choice
- Fluoroquinolones
- Third generation cephalosporins (e.g. ceftriaxone)
- Doxycycline
Vibrio vulnificus is a _____ fermenter.
Lactose
Can V. cholerae ferment sucrose?
Yes
Which Vibrio spp. is the least pathogenic for humans?
Vibrio alginolyticus
A strict halophile, requiring at least 1% NaCl.
Vibrio alginolyticus
V. alginolyticus NaCl requirement
at least 1%
V. alginolyticus can tolerate up to ______ NaCl.
10%
V. alginolyticus is associated with _________ infections.
wound, ear, and eye infections
______ is an occupational hazard with people in constant contact with seawater.
Vibrio alginolyticus
Specimens for Vibrio spp.
- Stool
- Wounds
Vibrio spp.
_____ specimens should be inoculated 2-4 hours within of collection onto appropriate agar media.
Stool
Vibrio spp.
Stool specimens should be inoculated within ________ of collection onto appropriate agar media.
2 - 4 hours
Vibrio spp.
Wounds are collected via _______ or _______.
Aspirates or Swab
Vibrio spp.
________ are collected via aspirates or swab.
Wounds
Vibrio spp.
For delayed processing of specimen…
The specimen is mixed in Cary-Blair transport medium and is refrigerated.
Which transport medium is used for delayed processing?
Cary-Blair
_______ is not recommended as a transport or holding medium for Vibrio spp.
Buffered Glycerol Saline
Why is buffered glycerol saline not recommended for Vibrio spp.?
Glycerol is toxic for Vibrio spp.
What is the gram stain reaction and morphology of the bacteria commonly seen in stool specimens?
Gram (-) bacilli
Which medium is used if there is a suspected presence of Vibrio spp.?
Alkaline Peptone Water (APW)
Why is APW used for Vibrio spp.?
It has some characteristics that favor the growth of Vibrio spp.
High pH and and NaCl content
Vibrio spp.
Time of Collection
As Soon As Possible (ASAP)
Vibrio spp.
Direct Microscopic Examination
- Gram (-) bacilli
- “shooting star” motility
Describe the motility of Vibrio spp.
“shooting star”
Which organism exhibits a “shooting star” motility?
Vibrio spp.
Which type of microscope is used to view the motility of Vibrio spp.?
Darkfield or Phase-contrast Microscope
Vibrio spp.
Culture Media
- Nutrient Agar or SBA (0.5% NaCl)
- MacConkey Agar (V. vulnificus)
- Thiosulfate, Citrate, Bile Salt Sucrose Agar (TCBS)
- Alkaline Peptone Broth or Water (1% NaCl and pH 8.5)
- Taurocholate Peptone Broth (pH 8.0 - 9.0)
- CHROMagar Vibrio
Vibrio spp.
Nutrient Agar and SBA content
0.5% NaCl
Thiosulfate, Citrate, Bile Salt Sucrose Agar
Differential Component
Sucrose
Thiosulfate, Citrate, Bile Salt Sucrose Agar
(-) Result
Green
Thiosulfate, Citrate, Bile Salt Sucrose Agar
Selective Component
Sodium Thiosulfate, Citrate, Bile Salt (oxgal), high pH
MacConkey Agar
(+) Result
Pink Colonies
MacConkey Agar
(-) Result
Colorless
Vibrio spp.
Alkaline Peptone Broth or Water content
1% NaCl + pH 8.5
Vibrio spp.
Taurocholate Peptone Broth content
pH 8.0 - 9.0
CHROMagar Vibrio is suited for _______.
- V. cholerae
- V. parahaemolyticus
- V. vulnificus
Vibrio spp.
Sucrose Fermenters (yellow colonies in TCBS)
- V. cholerae
- V. alginolyticus
- V. fluvialis
- V. furnassii
- V. cincinnatiensis
- V. metschnikovii
- some V. vulnificus
Vibrio spp.
Non-Sucrose Fermenters (green colonies in TCBS)
- V. mimicus
- V. parahaemolyticus
- P. damselae
- most V. vulnificus
Which Vibrio sp. does not grow on TCBS?
Grimontia (Vibrio) hollisae
Vibrio spp.
Colonial Appearance
Medium to large, smooth, opaque, iridescent with a greenish hue
Which organism?
Medium to large, smooth, opaque, iridescent with a greenish hue colonies
Vibrio spp.
Vibrio spp.
Beta-hemolytic NLF
- V. cholerae
- V. fluvialis
- V. mimicus
Vibrio spp.
Identification
- Oxidase Test
- Nutrient Broth w/ 6% salt
- String Test
- Vibrio static test O/129
ALL Vibrio spp. are oxidase (+) EXCEPT _______.
V. metschnikovii
ALL Vibrio spp. are indole (+) EXCEPT _______.
P. damselae
ALL Vibrio spp. are not capable of producing gas from glucose EXCEPT _______.
V. furnissii
ALL Vibrio spp. are lysine decarboxylase (+) EXCEPT _________.
- G. hollisae
- V. fluvialis
- V. furnissii
- V. vulnificus
ALL Vibrio spp. are arginine dihydrolase (-) EXCEPT _________.
- P. damselae
- V. fluvialis
- V. furnissii
ALL Vibrio spp. are ornithine decarboxylase (-) EXCEPT _________.
- V. cholerae
- V. mimicus
- V. parahaemolyticus
- V. vulnificus
Aeromonas spp.
Oxidase
Positive (+)
Aeromonas spp. are _______ fermenters.
Glucose
Aeromonas spp. resemble morphologically the family _________.
Enterobacteriaceae
How are Aeromonas spp. different from Enterobacteriaceae?
- Oxidase (+)
- Non-enteric gastrointestinal bacteria
Enterobacteriaceae is the opposite
A member of the Family Enterobacteriaceae, but is oxidase positive (+)
Plesiomonas spp.
Aeromonas spp. grow well on _______.
Blood Agar
Aeromonas spp.
Hemolytic Pattern
β-hemolytic
It causes gastroenteritis, which ranges from watery diarrhea to dysentery-like illness.
Aeromonas caviae
Dysentery means ______.
Presence of blood, mucus, and pus
It causes wound infections, and traumatic soft injuries (commonly described in patients with hematologic malignancies and/or liver disease).
Aeromonas hydrophila
Aeromonas spp.
Treatment of Choice
- Third generation cephalosporins
- Aztreonam
- Carbapenems
- Aminoglycosides
Aeromonas spp.
Intestinal Infections
- Acute secretory diarrhea
- Acute dysenteric form of diarrhea
- Chronic diarrhea
- Cholera-like disease
- Nebulous syndrome (traveller’s diarrhea)
Dysenteric illnesses are usually caused by _______.
Shigella spp.
Chronic diarrhea caused by Aeromonas spp. usually lasts for _______.
more than 10 days
Nebulous Syndrome is also known as _______.
Traveller’s Diarrhea
It is associated with Hemolytic-Uremic Syndrome (HUS) or kidney diseases that might require kidney transplantation.
Aeromonas veronii biovars. sobria and veronii
It is linked to cholera-like diseases characterized by abdominal pain, fever, and nausea.
Aeromonas veronii biovar. sobria
Which Aeromonas spp. often exhibit a β-hemolytic pattern?
- A. hydrophila
- A. veronii biovar. sobria
- A. jandaei
Aeromonas spp.
Culture
- SBA (β-hemolytic)
- MAC
- Modified Cefsulodin Irgasan-Novobiocin II Plate (CIN)
- CIN Medium
Can Aeromonas spp. ferment lactose?
Yes
Modified Cefsulodin Irgasan-Novobiocin II Plate (CIN) contents
- 4 ug cefsulodin
- Ampicillin
CIN Medium contents
15 ug cefsulodin
Appearance of Aeromonas spp. in CIN medium
Pink centered colonies (mannitol fermentation) with an uneven, clear apron resembling Yersinia enterocolitica
Aeromonas spp.
Spot Indole Test Result
Positive (+)
Aeromonas spp.
String Test Result
Negative (-)
Aeromonas spp.
Presumptive Identification
- Oxidase Test (+)
- Spot Indole Test (+)
- 0.5% NaCl
- String Test (-)
Similarities of Vibrio spp., Aeromonas spp., and Plesiomonas spp.
- Gram (-) Curved Bacilli
- Oxidase (+)
- Motile (+)
- Non-Enteric Bacilli (gastroenteritis)
Vibrio spp., Aeromonas spp., and Plesiomonas spp.
O129 Susceptibility Test
Susceptible: Vibrio spp.,
Resistant: Plesiomonas spp.Aeromonas spp.,
Vibrio spp., Aeromonas spp., and Plesiomonas spp.
Inositol Fermentation
To differentiate Vibrio spp. from Plesiomonas spp.
(+): Plesiomonas spp.
(-): Vibrio spp.
Vibrio spp., Aeromonas spp., and Plesiomonas spp.
Indole
To differentiate Aeromonas spp.
(+): Aeromonas spp.
(-): Vibrio spp., Plesiomonas spp.
Vibrio spp., Aeromonas spp., and Plesiomonas spp.
Mannitol Fermentation
(+): Vibrio spp.
(-): Aeromonas spp., Plesiomonas spp.
Vibrio spp., Aeromonas spp., and Plesiomonas spp.
Glucose Fermentation
(+): Vibrio spp., Aeromonas spp., and Plesiomonas spp.
(-): -
Vibrio spp., Aeromonas spp., and Plesiomonas spp.
6.5% NaCl
(+): Vibrio spp.
(-): Aeromonas spp., and Plesiomonas spp.
Vibrio spp., Aeromonas spp., and Plesiomonas spp.
TCBS
(+): Vibrio spp.
(-): Aeromonas spp., and Plesiomonas spp.
Vibrio spp., Aeromonas spp., and Plesiomonas spp.
Gelatin Liquefaction
(+): Vibrio spp., Aeromonas spp.
(-): Plesiomonas spp.
Campylobacter spp.
Aerotolerance
Microaerophilic
Requires reduced amount of oxygen
Specific shape of Campylobacter spp.
- Curved, comma-shaped, or S-shaped
- seagull wing shape
An organism characterized by a seagull wing shape.
Campylobacter spp.
A zoonotic organism known to cause abortion in domestic animals.
Campylobacter spp.
Campylobacter spp. is known to be associated with what syndrome?
Guillain-Barré Syndrome (GBS)
Patients suffering from GBS often test positive for _______.
Campylobacter antibodies
The most common cause of bacterial gastroenteritis.
Campylobacter jejuni
________ colonizes the stomach and cause inflammatory process until it develops gastritis which leads to ulcer.
Helicobacter pylori
It causes cramps and bloody diarrhea.
Campylobacter jejuni
A very common cause of diarrhea in humans.
Campylobacter jejuni
It has been isolated most frequently from blood cultures.
Campylobacter fetus subsp. fetus
A Campylobacter sp. that is rarely associated with gastrointestinal illness.
Campylobacter fetus subsp. fetus
Helicobacter spp.
Aerotolerance
Microaerophiles
_______ are not commonly isolated from culture media.
Helicobacter spp.
It is strongly associated with gastric, peptic, and duodenal ulcers as well as gastrointestinal carcinoma.
Helicobacter pylori
H. pylori is the major cause of ________.
Type B Gastritis
Type B Gastritis is caused by ________.
Helicobacter pylori
______ is associated with human gastroenteritis, generally in ICP.
Helicobacter cinaedi and Helicobacter fennelliae
Helicobacter spp. also associated with gastroenteritis
- H. canadensis
- H. canis
- H. pullorum
- H. winghamensis
An organism that is classified as a carcinogen.
Helicobacter pylori
Campylobacter spp.
Specimen Collection
- Stool samples (gastroenteritis)
- Rectal swabs (less preferred)
Both are transported using Cary-Blair medium
Campylobacter spp.
In collecting specimens, _______ are less preffered.
Rectal swabs
Helicobacter pylori
Specimen Collection
Gastric biopsy in Cysteine-Brucella Broth
Transported via: Stuart’s Medium
Cysteine-Brucella Broth contents
- 20% glycerol (frozen at -70 C)
- 10% sheep RBCs
- Vancomycin
- Trimethoprim
- Polymixin B
- Amphotericin B
- Cefalotin
Campylobacter spp. and Helicobacter spp.
Culture
- Campylobacter Blood Agar/ Campy Blood Agar
- Butzler’s Medium
- Skirrow’s Medium
- Campy-CVA (Cefoperazone, Vancomycin, Amphotericin B)
- Charcoal Cefoperazone Deoxycholate Agar (CCDA)
- Charcoal-based Selective Medium (CSM)
It provides better suppression to fecal normal flora.
Campy-CVA Medium
Campy-CVA medium is incubated at ________ for the recovery of Campylobacter spp.
37 C
Campy-CVA is a subculture for ______.
- Campylobacter fetus subsp. fetus
- Campylobacter fetus subsp. rectus
- Campylobacter fetus subsp. curvus
Campylobacter spp.
Incubation Period
- 42 C- majority
- 37 C - Campylobacter fetus subsp. fetus
Campylobacter spp.
Atmospheric Requirements
- 5% O2
- 10% CO2
- 85% N2
Helicobacter spp.
Atmospheric Requirements
- 5% - 10% O2
- 5% - 12% CO2
Campylobacter spp. and Helicobacter spp.
Colonial Morphology
Moist, runny-looking and spreading, non-hemolytic, round and raised, and others may be flat.
Campylobater fetus subsp. fetus
Colonial Morphology
Smooth, convex, transluscent
_________ can produce a dirty yellow pigment.
Campylobacter mucosalis and Campylobacter hyointestinalis
Campylobacter spp. and Helicobacter spp.
Microscopic Morphology
Curved, asporogenous, gram (-) bacilli/rods
Campylobacter spp.
Microscopic Morphology
- Long spirals or S- or seagull-wing shape
- “darting” motility via phase-contrast microscope using hanging operation
Campylobacter spp.
Oxidase Test Result
Positive (+)
Helicobacter spp.
Oxidase Test Result
Positive (+)
Campylobacter jejuni
Hippurate Hydrolysis Test
Positive (+)
Campylobacter jejuni is positive (+) in ____________.
Hippurate Hydrolysis Test
______ is capable of rapid Urease production.
Helicobacter pylori
Which culture medium is used for rapid urease production?
Christensen’s Urea Medium
Incubated at 37 C for 2 hours
Helicobacter pylori
Definitive Identification
- Rapid Urease Production
- Urea Breath Test
Explain the process of Urea Breath Test
Selective media for the cultivation of Campylobacter spp.
- Campy Blood Agar Plate
- Skirrow’s Medium
- Butzler’s Medium
- CCDA
Campy Blood Agar
Base
- Brucella Agar
- 10% sheep RBCs
Campy Blood Agar
Antimicrobial Agents
MNEMONIC: VTPAC
* Vancomycin
* Trimethoprim
* Polymixin B
* Amphotericin B
* Cefalotin
Skirrow’s Medium
Base
- Oxoid Agar Base
- Lysed, defibrinated horse RBCs
Skirrow’s Medium
Antimicrobial Agents
MNEMONIC: VTP
* Vancomycin
* Trimethoprim
* Polymixin B
Butzler’s Medium
Base
- Thioglycolate fluid with agar added
- 10% sheep RBCs
Butzler’s Medium
Antimicrobial Agents
MNEMONIC: BNACC
* Bacitracin
* Novobiocin
* Actidione
* Colistin
* Cefazolin
Charcoal Cefoperazone Deoxycholate Agar (CCDA)
Base
- Nutrient Agar
- Sodium Deoxycholate
- Charcoal
Charcoal Cefoperazone Deoxycholate Agar (CCDA)
Antimicrobial Agents
- Cefoperazone
- Amphotericin B