GROUP 5: 3GMT

Matawaran, Luis Paolo Montalbo, Carlo

Mercado, Coleen Iris Navarro, Francis

Miranda, Renz Osea, Rachelle

Family Vibrionaceae:
I. Vibrio spp.
General Characteristics:

 Grows well at very high pH (8.5-9.8)

 Rapidly killed by acid
 Grows well at 37ºC
 Ferments sucrose and mannose (cannot ferment arabinose and inositol)
 Halophiles
 Can grow on media containing 6% NaCl

Virulence Factors:

 Entero toxin – cholera toxin or choleragen

 Ability to induce hypersecretion of electrolytes (Na +, K+, HCO3-)

Microscopic morphology:

 Gram negative rods

 Pleomorphic under suboptimal conditions
 Comma shaped or curved bacilli (only in the initial gram stain of the clinical specimen)
 Usually appear as small , straight rods (on prolonged cultivation)
 Asporogenous
 05-08 µm x 1.5-3.0 µm

Colony morphology:

Vibrio cholorae Large yellow colonies in TCBS, Opaque and

granular in transmitted light

Vibrio parahaemolyticus Beta hemolytic in BAP, green colonies (TCBS)

Vibrio vulnificus Blue-green colonies in TCBS

Vibrio alginolyticus Large yellow colonies in TCBS

Culture Media:
  SBA (Sheep Blood Agar)
 Chocolate agar - large colonies that appear smooth, opaque and iridescent with greenish hue.
 Mac -Conkey agar - pathogenic vibrios usually grow as nonlactose fermenters
 Thiosulfate Citrate Bile Salts sucrose agar (TCBS) - Selective and differential. Differentiates sucrose-fermenting
species (yellow colonies) from nonsucrose-fermenting species (green colonies)

Biochemical Tests:

GROUP 1 GROUP 2 GROUP 3 GROUP 4 GROUP 5 GROUP 6

V. V. V. V. V. V. V. V. V. V.
choler mimic metschniiko cincinnatien damsel fluviali alginolytic parahaemolytic vulnific harver
TESTS ae us vii sis V. hollisae a s us us us yi
Nutrient
broth w/o
Nacl + + - - - - - - - - -
Nutrient
broth w/ 1%
Nacl + + + + + + + + + + +
Oxidase test     - + + + + + + + +
Nitrate
reduction
test     - + + + + + + + +
Myo-Inositol
Fermentatio
n     V + - - - - - - -
Arginine
dihydrolase         - + + - - - -
Lysine
decarboxyla
se         - V - + + + +
Ornithine
decarboxyla
se         -            

Pathogenesis:

 Most common in the clinical laboratory

1. V. cholerae (serogroups O1 and non-O1)
2. V. parahaemolyticus
3. V. vulnificus
4. V. alginolyticus

1) Vibrio cholera - Patients with infection experience a “rice water” stool sample. May experience:

 Dehydration
 Hypovolemic shock
 Metablic acidosis
 Rapid fluid & electrolyte loss
 Death

2)V. parhaemolyticus

 Self limited gastrointestinal diseases

 Kanawaga phenomenon – hemolysin and virulence
3)V. vulnificus

 Primary septicemia
 Wound infection

4)V. Alginolyticus – least pathogenic

II. Plesiomonas spp.

General Characteristics:

 Gram-negative comma bacilli

 Polarly flagellated pathogenic bacterium
 Motile
 Habitat is fresh water
 Oxidase positive
 Facultative anaerobe
 Glucose fermenter

Virulence Factors:

 Endotoxin production
 Beta-hemolytic
 Cholera-like enterotoxin production
 Large plasmid for invasion

Colony Morphology

 Grows readily on most routine media

 Shiny, opaque, slightly raised center, smooth
 Late-lactose fermenters
 Non-hemolytic
 Grows on most media routinely used in the clinical laboratory
 18-24 hours of incubation at 35°C, shiny, opaque, nonhemolytic colonies appear.
 Can grow well on CIN (cefsulodin-irgasin-novobiocin) as opaque colonies with an opaque apron.

Pathogenesis:

1) Plesiomonas Intestinal

 Gastroenteris
 Watery secretory diarrhea
 Colitis

2) Plesiomonas Extraintestinal

 Veterinarians and like jobs are more prone

 Bacteremia
 Meningitis
  Biliary tract disease

Family Campylobacteraceae:
I. Campylobacter spp.
General Characteristics:

 Gram negative, slim, motile and spirally curved rods

 All are Oxidase positive
 All are asaccharolytic
 All are strict anaerobes or microaerophilic
 Each cell has one polar flagellum
 Recognized world-wide as an important food borne pathogen

Virulence Factors:

 Motility and Adhesion - One of the most important aspects of virulences in Campylobacter is its nature of
interaction with intestinal cell lines
 Flagellum (protein: flagellin)
 Chemotaxis - important for intestinal colonization
 Toxin production : Enterotoxins and Cytotoxins

Colony Morphology:

 Moist or ‘runny-looking’
 Spreading
 Usually non-hemolytic
 Some are round and raised others are flat
 Tend to be colorless or gray
 Some produce tan or slightly pink coloration
 Does not ferment carbohydrates
 As moisture decreases colonies may form round convex glistening colonies with minimal spreading

 *Campylobacter mucosalis and Campylobacter hyointestinalis can produce a dirty yellow pigment

Campylobacter jejuni Moist, spreading, runny looking, non hemolytic, ‘runny-

looking’, round and raised others are flat

Campylobacter fetus subsp. fetus Smooth, convex, translucent

Campylobacter mucosalis and Campylobacter hyointestinalis Dirty yellow colonies

Microscopic morphology:

 Curved
 Non-spore forming
  Gram negative rods
 0.2-0.9 µm x 0.5-5.0 µm
 Single polar flagellum
 Enteric campylobacter
 as long as spirals
 S-shape
 Seagull-wing
 Microarophilic (<5%)

 May appear as coccobacilli (from older cultures or when exposed to air)

 ‘Darting-motility’ on hanging drop

 Stains poorly with gram stain

 Carbolfuchsin is the recommended counter stain

 If sarfranin is to be used, counterstaining should be extended for 2 to 3 minutes

 *Arcobacter spp. have a similar microscopic appearance to Campylobacter spp.

 *Helicobacter pylori may appear similar to Campylobacter spp. But Helicobacter spp. Have multiple flagella at
one pole

Culture Media:

Medium Base Antimicrobial agent

CAMPY blood agar plate Brucella agar Vancomycin

10% sheep red blood cells Trimethoprim
Polymyxin B
Amphotericin B
Cephalothin

Skirrow’s Oxoid blood agar Vancomycin

Lysed, defibrinated horse red blood Trimethoprim
cells Polymyxin B

Butzler Thioglycolate fluid with agar added Bacitracin

10% sheep red blood cells Novobiocin
Actidione
Colistin
Cefazolin

CCDA ( Charcoal cefoperazone Nutrient agar Cefoperazone

deoxycholate agar) Charcoal Amphotericin B
Sodium deoxycholate
Biochemical Tests:

Catalase Nitrate Urease H2 Hippurate Indoxyl 15 Growt 42 Nalidixi Cephalothin

Reduction S Hydrolysi Acetate C h C c Acid
s Hydrolysi 25 C
s
 C. Jejuni + + - - + + - - + + -
subsp.
Jejuni

C. jejuni V - - - V + - - - + +
subsp.
doylei

C. coli + + - V - + - - + + -

C. lari + + - - - - - - + - -

C. fetus + + - - - - - + - - +

C. + + - + - - - + + - +
hyointestinalis

C. upsaliensis - + - - - + - - + + +

C. concisus - + - V - - - - + - -

C. curvus - + - V V V - - + + ND

C. rectus V + - - V V - - W + ND

Pathogenesis:

 Abdominal pain
 Cramps
 Bloody diarrhea
 Fever and chills
 Nausea and vomiting
 Guillain-Barré syndrome – associated with infection; due to cross reaction with nerve cells in an
autoimmune response. Weakness of lower limbs progress in ascending manner.

II. Helicobacter spp.

 Helical, curved or straight bacilli  1-2 mm in diameter
 0.5-1.0μm x 2.5-5.0 μm  Translucent, convex colonies
 Lophotrichous  Slight hemolysis
 Appear coccoid in older cultures  Optimal growth temperature is at 35-37°C
 Microaerophiles
Pathogenesis:

1) Helicobacter pylori – recognized by immune system but Ab produced are not protective.

 Gastric infection
 Low-grade inflammatory process
 Chronic superficial gastritits
 Type B gastritis – stomach lining inflammed
 Associated with gastric carcinoma

Family Aeromonadaceae
I. Aeromonas spp.
 All are Oxidase positive  Motile
 Glucose fermenters  Gram (-) straight rods,
 Habitat are aquatic environments  Single polar flagellum
 Gram negative bacilli  1.0-3.5μm x 0.3-1.0μm
 Grow well on MacConkey agar  asporogenous
 Facultative anaerobe

Virulence Factors:

 Endotoxin production
 Enterotoxin production
 Vero cell cytotoxin
 Hemolysin
 Cytotoxic toxin

Colony morphology:

 Grows readily on most differential media used for enteric bacteria

 Large, round, raised, opaque colonies
 Smooth mucoid surface
 Hemolysis varies on SBA
o But most clinical species are strongly beta hemolytic except for Aeromonas caviae (which is nearly
always non-hemolytic or alpha-hemolytic at best)
 Pigmentation (translucent and white to buff colored)
 No growth on media containing 6% NaCl
 Some are lactose fermenters (ex. A.caviae)
 Appear as large round, raised, opaque colonies.
 Sheep blood agar
 Most major clinical species, such as A. hydrophila, A. veronii biovar sobria, and A. jandaei display strong β-
hemolysis.
 Aeromonas hydrophila, Strongly beta-hemolytic
Aeromonas veroniibiovar sobria, Aeromonas
jandaei
Aeromonas caviae Non-hemolytic or alpha-hemolytic

Biochemical Tests: (for differentiation of Aeromonas from Helicobacter)

Catalas Nitrate Ureas H2 Hippurate Indoxyl 1 Growt 4 Nalidixi Cephalothi

e Reductio e S Hydrolysi Acetate 5 h 2 c Acid n
n s Hydrolysi C 25 C C
s

A. butzleri -W + - - - + + + V V -

H. pylori + V + - - - - - V - +

H. + - - - - + - - - + +
fennelliae
H. cinaedi + + - - - V - - - + +

Pathogenesis:

1)Aeromonas Intestinal Infections

 Enteric Pathogen like Shigella, Salmonella and V. cholerae

 Several forms of diarrhea
 Cholera-like disease
 Traveller’s diarrhea

Most are self limiting but in the extremes of age and immunocompromised it is advisable to take meds.

2) Aeromonas Extraintestinal Infections

 Septiciemia
 Meningitis
 Wound infection
 Some sp. are invasive to liver, GI tract, respiratory tract and billiary ducts.