Emmanuel John A.

Genio
BSMT L3

GENUS VIBRIO
● Commonly found in a wide variety of aquatic environments, fresh, estuarine and marine or saltwater, BRACKISH WATER - primary habitat
● Gram (-), small, straight or slightly curved rod
● Robert Koch named it as Comma Bacillus
● BIOCHEMICAL PROPERTIES:
○ Facultative anaerobe
○ Ferments glucose
○ Oxidase (+)
○ Can reduce nitrate to nitrite except for Vibrio metschnikovii
● Dark Field microscopy:
○ “Darting Motility” or “Shooting Star Motility”
○ Polar flagella in broth
○ Peritrichous in solid media
● String Test (+)
● Mucoid stringing reaction after emulsification of colonies in 0.5% Sodium Deoxycholate
● Halophilic or salt-loving - Require addition of Sodium for growth and accurate identification except for V. cholerae and V. mimicus
● Not a normal flora
● MOT:
○ Ingestion of contaminated water, produce, meat, dairy products, or seafood
○ Exposure of disrupted skin and mucosal surfaces to contaminated water

LABORATORY DIAGNOSIS

Transport medium Gram Stain Biochemical test reactions Bacteriolysis Direct fluorescent Slide Dark Field Immobilization Grieg Test
antibody technique Agglutination Microscopy Test
(FAT) test

Cary and Blair
Buffered glycerol saline is
not recommended
(Glycerol is toxic for
Vibrios )
Gram (-) straight or Cholera Red Test / Nitroso Based on the lysis of “Darting Motility” or Used to demonstrate the soluble
slightly curved rods Indole Reaction: V. cholerae when “Shooting Star hemolysin of Vibrio cholerae
(comma-shaped ● A 24-hour culture inoculated to an Motility” biotype El tor
bacilli) grown in alkaline immuned guinea pig
peptone with NO3 and Uses sheep’s RBC + organism
tryptophan Pfeiffer’s incubated at 30°C - 37°C
● Addition of Phenomenon resulting to lysis
concentrated Sulfuric
Acid, H4 SO4 , :(+)
result is red color
indicating nitroso
indole

CULTURAL CHARACTERISTICS

BAP / CAP MCA TCBS Alkaline Peptone Water Tellurite Taurocholate Gelatin
Agar (TTGA)

Resembles members of NLF except V. vulnificus which maybe LF ● Medium of Choice Enrichment Broth with pH 8.4
the Enterobacteriaceae ● Thiosulfate Citrate Bile Salts Sucrose Agar
family ● Contains 1% NaCl After inoculation, the broth is incubated for
 Emmanuel John A. Genio
BSMT L3

● Bile Salts inhibit the growth of gram (+) organisms 5 – 8 hours at 35°C and sub cultured to
● Sucrose is for differentiation of the various Vibrio TCBS
species
● Bromothymol Blue and Thymol blue as pH indicator
● High pH I (8.4) inhibits the growth of other intestinal flora
● Those that ferment sucrose produces yellow colonies
and those that do not ferment sucrose produces green
colonies

VIBRIO SPECIES
Vibrio species Other significant VIrulence factor THREE MAJOR SUB GROUPS OF VIBRIO CHOLERAE
characteristics

Vibrio cholerae A notorious ● Cholera Toxin (CT) Vibrio cholerae Vibrio cholerae O1 Vibrio cholerae non-O1
pathogen ○ Powerful enterotoxin, a choleragen O139
○ Primarily responsible for cholera
○ Causes mucosal cells to hypersecrete water and electrolytes into Involved in Involved in Cholera epidemics Involved in Non-epidemic diarrhea
the lumen of GIT: Cholera epidemics and Extraintestinal
■ Profuse watery diarrhea leading to dramatic fluid loss Causative agent of Cholera / Asiatic
resulting in severe dehydration and hypotension that 8th Cholera Cholera/ Epidemic Cholera Strains resemble V. cholerae but
without medical intervention, frequently lead to DEATH epidemic due to a fail to agglutinate O1 antisera
■ RICE-WATERY STOOL” – hallmark of cholera toxin new serogroup All strains share a common Flagellar
activity recently emerged in Antigen (H Ag) Phenotypically similar to toxigenic
■ 1 liter / hour India V. cholerae O1 but most lack
● Zonula Occludens Toxin (Zot) On the basis of their Somatic Antigen (O cholera toxin gene and appear
○ Enterotoxin Found in chromosomes (for Enterobacteriaceae in Ag), it is divided into six (6) serogroups to cause a milder form of
plasmids gastroenteritis or cholera-like
○ Hypersecretion of water and chloride into the intestinal disease
lumen and inhibits absorption of Sodium ions
○ Results to diarrhea and profound dehydration, altering Have also been implicated in
acid-base balance extraintestinal infections such as
○ May lead to shock followed by death after 18 hours if not treated cholecystitis,
immediately
● Accessory Cholera Enterotoxin (Ace) Ear infections, cellulitis and
● O1 and O139 Somatic Antigens septicemia
● Hemolysins / Cytotoxins
● Motility
● Chemotaxis
● Mucinase Production
● Toxin Coregulated (TCP)
○ Provides the means by which bacilli attach to mucosal cells for
the release of cholera toxin

Other significant characteristics Associated infection Growth medium Biochemical test reaction

Vibrio Second most common Vibrio species implicated in Number 1 cause of “Summer Diarrhea” in Most clinical strains produce a heat stable hemolysin that is (+):Hemolysis - Kanagawa strain
parahaemolyticus gastroenteritis Japan able to lyse human RBC in a special high salt mannitol
medium called Wagatsuma Agar
Self-limiting
 Emmanuel John A. Genio
BSMT L3

Found in aquatic environment but appears to be

limited in coastal or estuarine areas despite a
halophilic requirement for 1% – 8% NaCl

Associated with marine species, including oysters,

clams, crabs, lobsters, scallops, sardines and shrimps

Can also be serotyped by means of its O and K

Antigens
● Used only in outbreaks or in large
epidemiologic studies

Previously Pasteurella parahaemolyticus

Chromogenic Vibrio agar was developed for the

recovery of V. parahaemolyticus from seafood
● Also supports the growth of other Vibrio
species
● Colonies appear as white to pale blue and
violet

Vibrio vulnificus Found in marine environments Second most serious type of Vibrio– Commonly referred to as lactose
associated infections after cholera (+) Vibrio

Clinical Infections:
● Fatal Septicemia especially in patients
with underlying liver disease
● Wound Infections

VIbrio alginolyticus Inhabitant of marine environment Least pathogenic Strict halophile

Previously Vibrio parahaemolyticus biotype 2

OTHER VIBRIO SPECIES

● Vibrio mimicus (previously V. cholerae sucrose (-)

● Vibrio fluvialis
● Vibrio metschnikovii
● Vibrio cincinnatiensis
● Vibrio furnissil
● Vibrio harveyi (previously V. carchariae)
● Grimontia hollisae (previously V. hollisae
● Photobacterium damselae (previously V. damsela

Normally found in
Found generally in fresh
water but may also be found
in brackish water or marine
water with low salt content
Widely distributed in
freshwater, estuarine and
marine environments
Frequently isolated from
retail produce sources and
meat products
● Aeromonas caviae
● Aeromonas hydrophila
● Aeromonas sobria
● Aeromonas veronii
Emmanuel John A. Genio
BSMT L3
GENUS AEROMONAS
Other significant characteristics Associated infection Culture Characteristics Odor Biochemical reactions
Serotyping system is based on O Ag Can cause gastroenteritis most frequently Does not grow in 6% NaCl that fruity odor Oxidase (+)
in children but its role in intestinal infection is distinguishes them from Vibrio spp
Gram (-) straight rods not yet clear Catalase (+)
Growth temperature; 0 – 45°C
Most are motile with single polar Similar to Vibrio species in terms of type of Ferments glucose
flagellum (Monotrichous) infections they cause Facultative anaerobe
Biochemical reaction similar to
Responsible for disease syndrome among a Grows readily on conventional laboratory pseudomonas except mannitol
variety of warm blooded and cold blooded medium fermentation and gelatinase
animals including humans reaction:
Produces colonies resembling ● Pseudomonas:
Produces heat-labile and heat-stable Pseudomonas ○ Gelatinase (-)
enterotoxin ○ Non-Mannitol
Has a greenish ground glass Fermenter
CLINICAL INFECTIONS: appearance and gives off fruity odor ● Aeromonas:
● Gastroenteritis ○ Gelatinase (+)
● Wound Infection β-hemolytic on BAP ○ Mannitol
● Septicemia Fermenter
CINAgar
COMPLICATIONS: ● Cefsulodin-Irgasan-Novobiocin
● HUS
● Kidney Disease
AEROMONAS SPECIES
 Emmanuel John A. Genio
BSMT L3

GENUS CHROMOBACTERIUM
● Infections are not associated with GIT but acquired following contact of broken skin with contaminated soil or water that leads to fulminant, lifethreatening systemic infect
● Found in soil and water

Chromobacterium species Other significant characteristics Associated infection Culture Characteristics Biochemical reactions Odor

Chromobacterium violaceum Only species in the genus Cellulitis Pigmentation interferes in the oxidase Glucose fermenter Almond like odor
test
Most strains produces VIOLACEIN Abscess Formation ● Must be cultured anaerobically Indole (+)
● Ethanol – soluble violet pigment wherein in such condition does
Appears as black to purple colonies due Septic Shock not produce pigment Lysine (-)
to the production of the purple pigment ● Typical violet pigment is
Directed therapy is required enhanced optimally when Oxidase (+)
Often resistant to β-lactams and colistin cultures are incubated at room
temperature Ornithine (-)
A Polymerase Chain Reaction (PCR)
amplification assay has been developed for Β-hemolytic on BAP Does not ferment maltose and
its identification mannitol

GENUS CAMPYLOBACTER

● Gram (-) small curved rods showing S shaped, “seagull-wing” appearance

● Inhabitants of GIT and reproductive organs of animals
● Motile with single polar flagellum
● Exhibits a characteristic darting motility on hanging drop preparations
● Visualized under phase-contrast microscopy or dark-field microscopy
● Require selective media

Other significant characteristics Mode of transmission Associated infection Biochemical reactions

Assacharrolytic ● DIrect contact to infected animals such as dogs, Known to cause abortion in domestic animals such as cattle, Oxidase (+)
cats, and birds sheep and swine
Microaerophilic environment is required ( ● Indirectly via consumption of contaminated catalase (+)
5%-10% O ) water/dairy products or poultry Gastroenteritis associated with Campylobacter is usually self
● Person-person limiting
Inhabitants of GIT and reproductive organs of ● Sexually transmissible
animals
 Emmanuel John A. Genio
BSMT L3

LABORATORY DIAGNOSIS
SPECIMEN OF TRANSPORT DIRECT DETECTION ANTIGEN DETECTION MEDIA CULTIVATION IDENTIFICATION ANTIMICROBIAL
CHOICE SUSCEPTIBILITY TESTING
AND THERAPY

Feces / Stool Cary – Blair Medium or ● Gram Stain ● Several Campy-BAP STOOL ● Colonies are gray ● Not standardized
● Rectal Campy-Thio - if deley is ● PCR commercially ● Enriched selective BAP ● Inoculated in two to pinkish or ● Not routinely performed
Swab can anticipated in atleast 2 available Ag ● Brucella Agar Base, selective medium yellowish gray and ● Susceptible to many
also be hours detection systems Sheep RBC, ● Two sets should be slightly mucoid antimicrobial agents
accepted are available Vancomycin, incubated looking including macrolides,
● EIA (Enzyme Trimethoprim, Polymixin separately in 37°C ● Some colonies tetracyclines,
Blood Immunoassays) B, Amphotericin B and and 42°C may exhibit a aminoglycosides and
Cephalothin ● Extended tailing effect along quinolones
incubation up to 48 the streak line ● Erythromycin is the
Campy Medium (CVA) – 72 hours ● Wet method from drug of choice for
● Cefoperazone, broth: darting severe gastroenteritis
Vancomycin and BLOOD motility ● Ciprofloxacin as
Amphotericin B ● Organism is ● Gram Stain: alternative therapeutic
capable of growth curved gram (-) option
Campylobacter agar base in less than 5 rod ● Previously,
blood free (CCDA) days in most blood Fluoroquinolones were
● Modified agar without culture media used however,
blood ● Extended increasing
● Blood is replaced with incubation period Fluoroquinolone
charcoal, Sodium is required for resistant strain
pyruvate and Ferrous better recovery up worldwide had been
sulfate to 2 weeks identified
● Subcultures ● Parenteral therapy is
Skirrow’s Medium should be used to treat systemic
● Vancomycin and incubated in 5 % - infections
Polymixin B 10% O
(microaerobic
environment)
● Turbidity may not
be visible in blood
culture, blind
subculture may be
necessary
● Microscopic
examination using
acridine orange can
also be done
● Presence in blood
is effectively
detected through
CO monitoring
● Isolation from
sources other than
blood and feces is
 Emmanuel John A. Genio
BSMT L3

extremely rare

CAMPYLOBACTER SPECIES
Campylobacter species Other significant characteristics Associated infection Growth requirement

Campylobacter jejuni Associated with warm-blooded animals primarily birds Most common cause of bacterial gastroenteritis worldwide Stool cultures are incubated at 42°C

Cramps and bloody diarrhea may follow the initial mild Ideal atmospheric environment contains a mixture of 5-10% O
abdominal pain and 10% CO 2 2

Campylobacter fetus spp fetus Isolated most frequently from blood cultures Rarely associated with gastrointestinal illness n/a

Blood is the specimen of choice for isolation Majority of infections occur among immunocompromised and
elderly patients
Associated with cattle and sheep

OTHER CAMPYLOBACTER SPECIES

● Campylobacter coli – associated with pigs

● Campylobacter concisus
● Campylobacter curvus
● Campylobacter fetus ssp. venerealis
● Campylobacter gracilis
● Campylobacter hyointestinalis ssp. hyointestinalis
● Campylobacter mucosalis
● Campylobacter jejuni ssp. doylei
● Campylobacter jejuni ssp. jejuni
● Campylobacter lari
● Campylobacter rectus
● Campylobacter showae
● Campylobacter sputorum biovar sputorum
● Campylobacter upsaliensis

ARCOBACTER
Other significant characteristics Associated disease

● Slow-growing ● Associated with gastroenteritis

● Fastidious
● Asaccharrolytic
● Found in the environment and in untreated water
● Also prevalent in commercially prepared meat including beef, chicken, pork, lamb and poultry
● Arcobacter cryaerophilus – formerly Campylobacter cryaerophila
● Arcobacter butzleri

Identification, Associated disease, Epidemiology MOT, lab diagnosis
● Curved, microaerophilic gram (-) rods
● Strongly associated with gastric and duodenal ulcers
● Now recognized as a major cause of chronic superficial gastritis or type B gastritis
○ A condition formerly associated with stress and chemical irritants
Helicobacter species Identification
Helicobacter pylory Spiral-shaped organisms resembling Campylobacter
spp.
ANTIMICROBIAL SENSITIVITY TESTING & THERAPY
Except for Metronidazole and Clarithromycin most
laboratory susceptibility assays are unsuccessful in
predicting clinical outcome
Recommended to use E-test and Agar or broth
Emmanuel John A. Genio
BSMT L3
GENUS HELICOBACTER
HELICOBACTER SPECIES
Mode of transmission Epidemiology Associated disease
● Oral – oral Isolated from human stomach in 1983 and was named Capable of colonizing the mucous layer
● Fecal – oral Campylobacter pyloridis then became Campylobacter pylori or of the antrum and fundus of the
● Mother – child was called as Campylobacter-like organism stomach
No human reservoir Genus Helicobacter was established in 1989, hence Once acquired, it colonizes the stomach
renamed as Helicobacter pylori for a long time and causes a low grade
Had been isolated from inflammatory process producing a
feces and dental plaque Antibody surveys indicate that approximately 50% of chronic superficial gastritis
adults >60 years old are infected by H. pylori
Speculation: long-term H. pylori
Gastritis incidence increase with age infection leading to chronic gastritis
is an important risk factor for gastric
Occasionally been cultured from feces and dental plaque carcinoma
Has been identified in more than 75% of gastric ulcer
patients
US: has been reported to colonize 20%-40% of the adult
population
Developing countries: Asia, Africa and South America the
incidence is reported to be as high as 80%-100% of the adult
population
Incidence is attributed to poor sanitary conditions
LABORATORY DIAGNOSIS
SPECIMEN IDENTIFICATION / BIOCHEMICAL TEST REACTION MEDIA
Gastric biopsy materials Microscopic morphology similar to Campylobacter but has Stuart Medium can be used to
multiple flagella at one pole maintain viability of organism if delay
is anticipated
Oxidase (+)
CAP and Skirrows Medium

dilution methods
Therapy is problematic: H. pylori readily becomes
resistant when Metronidazole, Clarithromycin,
Azithromycin, Rifampicin or Ciprofloxacin is
prescribed as a single agent
Current regimen recommend triple drug therapy
including Metronidazole, a bismuth salt , and either
amoxicillin or tetracycline
Alternative for Metronidazole resistant strains:
Omeprazole or Lansoprazole and amoxicillin or
Clarithromycin
Relapses occur often
Infections associated with enteritis and proctitis may
respond to quinolones, however appropriate therapy
has not been established
No vaccines available, but several vaccines are under
development
Helicobacter cinaedi formerly Campylobacter cinaedi
Helicobacter fennelliae formerly Campylobacter fennelliae
Emmanuel John A. Genio
BSMT L3
Catalase (+)
Must be incubated in an environment
Rapid Urease Reaction with increased humidity
● Gastric biopsy specimen may be placed onto a
Christensen’s Urea Medium and incubated for 2 hours,
rapid color change suggests the presence of H. pylori
Urea Breath Test
● Recommended for monitoring therapy
● Patient is given C-labeled urea to drink
● Urea is degraded by the urease activity of H. pylori
● 14CO2 is released and detected in the exhaled
breath by a scintillation counter