BACTE_Finals_Lec: Vibrio, Aeromonas, Camplylobacter (copy)

CM = Colony Morphology

 Vibrio species

Commonly found in wide variety of aquatic environments,  they are responsible for marine and freshwater diseases, including gastroenteritis.

V. cholerae

most common Vibrio species that causes diarrhea 

1. consumption of raw seafood

2. recent immigration of foreign travel

3. Gastroenteritis with cholera like or rice water stools

4. Accidental trauma with marine water products (shellfish,oyster or clam shells, fishhooks

Factors in identifying vibrio infection

1. V. cholerae O1

2. V. cholerae O139

3. V. parehaemolyticus

4. V vulnificus

most common vibrio spp.

Asporogenous, curved (comma) or pleomorphic gram (-) rods

Micro Morpho of Vibrio

V. metschnikovii

All vibrio spp. are CPOP and Nitrate reduction except what sp.

susceptible @ 150 ug

Vibrio are 0/129 (Susceptible/resistant), at what concencetration

MAC

> TCBS (Thiosulfate Citrate Bile Sucrose) Agar

Culture media where vibrio spp. can grow?

> what routine media?

> Vibrio

> 0.5% Sodium desoxycholate

these ____ spp. are (+) in string test after emulsification of colonies in _% _____

V. cholerae, V. mimicus

The only Vibrio spp. that are not halophilic (salt-loving)

sodium

What is added on the culture media in order for Vibrio spp. to grow?

1. V. cholerae O1

2. V. cholerae O139

3. V. cholerae non-O1

3 major subgroups of Vibrio

Flagellar (H) antigen

Somatic (O) Antigen

V.F of  3 major subgrps. of V. cholerae

Ogawa (India)

Inaba (Philippines)

Hikojima (Japan)

3 serotypes of V. Cholerae O1 organims

V. cholerae

Causative agent of cholera aka Asiatic cholera or epidemic cholera

Cholera toxin (Choleragen)

a protein enterotoxin that is responsible for rice watery stool

1. azithromycin (AZM)

2. ciprofloxacin (CPFX)

Antibiotics for Cholera

2 types of cholera

Classical type

El tor Type - more serious type

1. Voges-Proskauer Test

2. Polymyxin B

3. Serological test using Chicken RBC as Antigen

Tests used to differentiate Classical Type and El Tor Type of Cholera

<insert image for Classic vs El Tor Type>

Determine the test result

VIBRIO PARAHAEMOLYTICUS

• causes summer diarrhea

• raw, improperly cooked seafood (i.e. oysters)

• Kanagawa-toxin positive

Kanagawa-toxin

This is a toxin that Produces a heat-stable hemolysin that is able to lyse human erythrocytes in a special, high salt mannitol medium Wagatsuma Agar

Wagatsuma Agar

a special high salt mannitol medium used for the detection of Kanagawa Toxin

V. alginolyticus

(V. alginolyticus/V. vulnificus) is Least pathogenic; most frequently isolated

V. vulnificus

(V. alginolyticus/V. vulnificus)

has Two categories of infection:

1. primary septicemia

2. wound infections

Vibrio

This genus:

in SBA and CAP colonies: medium to large colonies that appear smooth, opaque, and iridescent with a greenish hue

a and B- hemolysis

hemolytic pattern of Vibrio on SBA

V. vulnificus

T/F

ALL Vibrio spp. in MAC are Lactose fermenter except what Vibrio sp.

No, V. vulnificus is NLF

Are all Vibrio spp. Lactose fermenters?

Vibrio spp.

Determine what genus:

MAC and CIN agar can give false-positive oxidase reactions

Cary-Blair Media

Transport media for Vibrio

not recommended

For Vibrio spp:

Buffered Glycerol Saline - (not recommended/recommended) for transport

TCBS (thiosulfate citrate bile salt sucrose agar)

Recommended selective medium for Vibrio spp.

Vibrio spp. - NON-SUCROSE FERMENTER

1. V. mimicus 

2. V. parahaemolyticus 

3. V. damsela 

4. V. vulnificus strains

(My Pair of Dancing Vulgar)

Vibrio spp. - SUCROSE FERMENTER

Tip: FFACC Me

1. V. cholerae

2. V. alginolyticus

3. V. fluvialis

4. V. furnissii

5. V. cincinnatiensis

6. V. metschnikovii

alkaline peptone water with 1% NaCl can be inoculated (at least 20 mL) and incubated for 5 to 8 hours at 35° C before subculturing in TCBS

Enrichment procedure for Vibrio spp. before subculture in TCBS

• V. cincinnatiensis

• some strains of V. metschnikovii

All Vibrio spp. are inositol nonfermenter except

(+) in String test

(+) in Vibriostatic agent O/129

what tests separates Vibrio from Aeromonas

V. cholerae - B-hemolysis

V. parahaemolyticus - a-hemolysis

Based on the picture describe the hemolytic pattern of the ff Vibrio spp.

(-) in inositol fermentation

what test separates Vibrio from Plesiomonas

(+) Oxidation rxn

what test separates Vibrio from Enterobacteriaceae excluding Plesiomonas

(+) Carbohydrate metabolism

what test separates Vibrio from Pseudomonas

Aeromonas

• these species live in similar environment like Vibrio but prefer fresh water

A. hydrophila

A. sobria

A. caviae

Major clinical specimens of Aeromonas

A. caviae

most commonly isolated Aeromonas sp.

Motile with single tuft of flagella

Aeromonas Motility:

A. caviae

Aeromonas sp. mostly associated with gastrointestinal (GI) infection

Cellulitis

most common presentation of wound infection by Aeromonas sp.

Aeromonas

Genus:

COLONY APPEARANCE: large, round, raised, opaque colonies with an entire edge and a smooth, often mucoid surface

strong B-hemolysis

hemolytic pattern of Aeromonas

Mannitol

Aeromonas spp:

CIN colony appearance – pink-centered colonies (____ fermentation)

Oxidase test

this test separates the Oxidase (+) Aeromonas from Oxidase (-) Yersinia

positive (+)

ALL AEROMONAS  are Indole (+/ -)

Aeromonas: (-)

Plesiomonas: (+)

Aeromonas vs. Plesiomonas in term of Inositol test

String test where Aero. are (-) and Vibrio is (+)

What test separates Aeromonas from Vibrio

A. caviae

Aeromonas sp. that is Lactose Fermenter

True

T/F:

Aeromonas are overlooked as normal biota E. coli in MAC

resistant

Aeromonas spp:

Vibriostatic O/129 (resistant/susceptible)

Camp. are asaccharolytic (non-fermenters)

what makes Campylobacter diff from Vibrio in terms of carb. fermentation

C. jejuni subsp. jejuni

C. coli

C. fetus subsp. fetus

3 spp. of Camp.

C. jejuni

among the Campylobacter spp.:

most common cause of bacterial gastroenteritis worldwide

C. fetus

Camp. sp.: causative agent of bacteremia

C. jejuni

Camp. sp.: causative agent of Guillain-Barre Syndrome

GUILLAIN- BARRE SYNDROME

an autoimmune disorder with acute paralysis due to PNS destruction caused by Campylobacter jejuni

Helicobacter pylori

Helicobacter s.: important species

H. pylori

bacteria that is a major cause of type B gastritis or peptic ulcer

Helicobacter pylori

_______ (bacteria) is a carcinogenic agent that causes gastric carcinoma during long term infection

True

T/F: Camp. and Hel. are considered fastidious

Cary-Blair medium

transport medium for Campy. spp.

Buffered-glycerol saline

transport medium that should be avoided in transporting Campy. spp.

blood with incubation at 35◦C to 37◦C

specimen choice for C. fetus

gastric biopsy material

Spx. for Helicobacter pylori

1. Stuart medium

2. Cysteine-brucella Broth with 20% glycerol and frozen at −70° C

1. Transport medium fro H. pylori specimens; and 

2. their Storage condition

CampyBAP (blood agar plate)

selective media for C. jejuni

Butzler medium and Skirrow’s medium

other medium for C. jejuni aside from Campy Blood Agar Plate (CampyBAP)

• CAP

• Brucella agar with 5% Horse red blood cells

Commonly used media for H. pylori

Skirrow’s agar

Selective media for H. pylori

Vancomycin

Trimethoprim

Polymyxin B

Amphotericin B

Cephalothin

Antimicrobial agents of Campy Blood agar plate

(Tip: VTPAC)

Incubation temp for

C. jejuni - 42 C

H. pylori - 37 C

Incubation temp for

C. jejuni

H. pylori

Both

(Both/Neither) Campylobacter spp. and Helicobacter spp. require a microaerophilic and capnophilic environment

aerobic (O2) and anaerobic (CO2) conditions for

Campy. - – 5% O2, 10% CO2 

Heli. - 5% to 10% O2 and 5% to 12% CO2

aerobic (O2) and anaerobic (CO2) conditions for

Campy. 

Heli.

Campylobacter spp.

• curved non–spore-forming, gram-negative rods

• May appear as long spirals or ‘S’ or seagull-wing shapes

Carbol fuchsin

recommended as a counterstain fro Campy.

Campylobacter

“Darting” motility (corkscrew movement) on hanging drop preparations

Brucella or Tryptic Soy Broth

(motility can be inhibited by Distilled water or saline)

medium used for motility testing of Campy. spp.

Helicobacter spp.

➢ Curved or U-shaped

➢ Motile with multiple flagella at one pole

smooth, convex, translucent colonies

CM of C. fetus subsp. fetus

moist, runny looking, and spreading

• nonhemolytic

• some are round and raised and others may be flat

CM of C. jejuni and other enteric Campylobacters

	Campy	Heli.
Oxidase	+	+
Urease	(-) (C. jejuni)	+ (H. pylori)
Hippurate Hydrolysis Test	(+) (C. jejuni)	-

	Campy	Heli.
Oxidase
Urease
Hippurate Hydrolysis Test