CURVED GRAM-NEGATIVE RODS

Vibrio spp.
Aeromonas
Helicobacter spp.
Campylobacters pp.
◦ Vibrios are small (0.5 to 1.5 to μm), curved, comma-shaped,
gram-negative rods
◦ They are facultatively anaerobic, fermentative rods
◦ Oxidase reactions positive
◦ Most vibrios have polar flagella
◦ The genus is composed of more than 200 species of curved rods
◦ Three species are particularly important human pathogens:
Vibrio cholerae - cholera
Vibrio parahaemolyticus - gastroenteritis, wound infection
Vibrio vulnificus - wound infection
◦ Vibrio species can grow on a variety of media within a broad
temperature range (from 14° C to 40° C):
- including blood agar and MacConkey agar

- thiosulfate citrate bile salts sucrose [TCBS] agar

- enrichment broth (alkaline peptone broth; pH 8.0-9.5)
◦ All species of Vibrio require sodium chloride (NaCl) for growth
(halophilic species)- except V. cholerae
◦ They are susceptible to stomach acids
◦ The O polysaccharide is used to subdivide Vibrio species into
serogroups
◦ Members of the O1 group cause epidemic disease, whereas
non-O1 group (02-0140) either cause sporadic disease or are
nonpathogens
◦ V. cholerae O1 organisms have three serotypes:
Inaba
Ogawa
Hikojima
◦ V. cholerae O1 and O139 produce cholera toxin
◦ V. cholerae O1 organisms have two biotypes:
V. cholerae biotype cholerae
V. cholerae biotype El Tor
◦ V. cholerae secretes an enterotoxin called choleragen (cholera toxin)
◦ Choleragen consists of an A (active) subunit and a B (binding) subunit
◦ The B subunit, which is a pentamer composed of five identical
proteins, binds to a ganglioside receptor on the surface of the
enterocyte
◦ The A subunit is inserted into the cytosol, where it catalyzes the
addition of ADP-ribose to the Gs protein
◦ The genes for cholera toxin and other virulence factors are carried on
a single-stranded DNA bacteriophage called CTX
◦ The pili that attach the organism to the gut mucosa are the receptors
for the phage
Virulence Factor Biologic Effect
Cholera toxin Hypersecretion of electrolytes and
water

Toxin co-regulated pilus Binding site for CTXΦ; mediates adherence to

intestinal mucosal cells

Chemotaxis protein Adhesin factor

Accessory cholera enterotoxin Increases intestinal

fluid secretion

Zonula occludens toxin Increases intestinal permeability

Neuraminidase Modifies cell surface to increases GM1

binding sites for cholera toxin
◦ V. cholerae is transmitted by fecal contamination of water and food,
primarily from human sources
◦ Human carriers are frequently asymptomatic and include individuals
who are either in the incubation period or convalescing
◦ The main animal reservoirs are marine shellfish, such as shrimp and
oysters
◦ Ingestion of these without adequate cooking can transmit the
disease
◦ Typical cholera has a rapid onset
◦ There are no red blood cells or white blood cells in the stool
◦ Rice-water stool is the term often applied to the nonbloody effluent
◦ There is no abdominal pain, and subsequent symptoms are referable to the
marked dehydration
◦ The patient is afebrile
◦ The loss of fluid and electrolytes leads to cardiac and renal failure
◦ No other disease produces dehydration as rapidly as cholera
◦ Acidosis and hypokalemia also occur as a result of loss of bicarbonate
◦ The mortality rate without treatment is 60%
◦ A bacteriologic diagnosis is accomplished by
isolation of V. cholerae from the stool
◦ For diagnosis of sporadic cases:
The organism grows on common clinical laboratory
media such as alkaline peptone broth (enrichment
broth- pH 8.6), blood agar and MacConkey agar, but
its isolation is enhanced by a selective medium
(thiosulfate–citrate–bile salt–sucrose - TCBS agar )
◦ The organism is readily identified by biochemical
reactions
◦ Presumptive diagnosis of V. cholerae can be confirmed by
agglutination of the organism by polyvalent O1 or non-O1 antiserum
◦ A retrospective diagnosis can be made serologically by detecting a rise
in antibody titer in acute- and convalescent-phase sera

◦ High sensitivity and specificity have been reported more recently

using polymerase chain reaction (PCR) assay and real-time nucleic acid
sequence-based amplification assays for detecting vibrios in stool and
environmental samples
◦ Oral or intravenous fluid and electrolyte replacement is crucial
◦ Antimicrobial therapy can reduce duration and severity
◦ A single dose of azithromycin, doxycycline or ciprofloxacin
◦ Water sanitation and cooking shellfish prevent infection
◦ Vaccines:
- prepared from whole cells

- CT B subunit have been disappointing, providing protection that is

not longlasting
◦ Current interest includeslive attenuated vaccine strains because of their
potential to stimulate the local sIgA immune response
◦ V. parahaemolyticus is a marine organism transmitted by ingestion of raw
or undercooked seafood, especially shellfish such as oysters
◦ The clinical picture caused by V. parahaemolyticus varies from mild to quite
severe watery diarrhea, nausea and vomiting, abdominal cramps, and fever
◦ The illness is self-limited, lasting about 3 days
◦ V. parahaemolyticus is distinguished from V. cholerae mainly on the basis
of growth in NaCl:
- V. parahaemolyticus grows in 8% NaCl solution, whereas V. cholerae
does not
- No specific treatment is indicated, because the disease is relatively
mild and self-limited
◦ Disease can be prevented by proper refrigeration and cooking of seafood
◦ V. parahaemolyticus produce a thermostable direct hemolysin (TDH; also
called Kanagawa hemolysin)
◦ V. vulnificus is also a marine organism
◦ Curved gram-negative rods, facultative anaerobic; require salt for growth
◦ Virulence associated with presence of polysaccharide capsule and
hydrolytic enzymes
◦ It causes severe skin and soft tissue infections (cellulitis), especially in
shellfish handlers, who often sustain skin wounds
◦ It can also cause a rapidly fatal septicemia in immunocompromised
people who have eaten raw shellfish containing the organism

◦ Culture wounds and blood

◦ The recommended treatment is doxycycline
◦ Aeromonas is a gram-negative, facultative anaerobic fermentative rod
◦ The organisms are ubiquitous in fresh and brackish water
◦ Thirty species and 12 subspecies of Aeromonas have been described
◦ The most important pathogens are:
- Aeromonas hydrophila
- Aeromonas caviae
- Aeromonas veronii biovar. sobria
 Aeromonas species cause three
forms of disease:

1.Diarrheal disease in otherwise

healthy people
2. Wound infections
3. Opportunistic systemic disease in
immunocompromised patients
(particularly those with hepatobiliary
disease or an underlying malignancy)
◦ Antimicrobial therapy is necessary in patients with chronic diarrheal
disease, wound infections, or systemic disease
◦ A fluoroquinolone can be used initially for empiric therapy

◦ But activity should be confirmed with in vitro susceptibility tests

◦ Plesiomonas are straight (0.8 to 1 μm by 3 μm), gram negative
bacilli that occur singly, in pairs, or in short chains or filamentous
forms
◦ They are motile by monotrichous or two to five lophotrichous
flagella
◦ The genera Plesiomonas and Shigella share both biochemical and
antigenic features
◦ P. shigelloides is oxidase-positive
◦ P. shigelloidesis found in both soil and aquatic environments, but
because of intolerance to increased NaCl and a minimum growth
temperature of 8° C

◦ They are widely distributed among both warm- and cold-blooded

animals, including dogs, cats, pigs, vultures, snakes, lizards, fish,
newts, and shellfish
Gastroenteritis
Three major clinical types of gastroenteritis are caused by Plesiomonas:
◦ The more common watery or secretory diarrhea
◦ A subacute or chronic disease that lasts between 14 days and 2 to 3 months
◦ A more invasive, dysenteric form that resembles colitis
Extraintestinal Infection
Serious infections, such as bacteremia and meningitis, usually occur only in
severely immunocompromised patients or neonates

◦ Penicillins combined with a β-lactamase inhibitor, as well as trimethoprim-

sulfamethoxazole, are active