GENIO, EMMANUEL JOHN A.

BSMT L3

GENUS CORYNEBACTERIUM
● Coryneforms or diphtheroids
● Found worldwide in fresh and saltwater, soil, and air
● Closely related to Mycobacteria and Nocardia: collectively referred to as CMN group (share a common cell wall structure)
● OTHER CORYNEBACTERIUM DIPHTHERIAE SUBSPECIES THAT PRODUCES DIPHTHERIA TOXIN: (only produces toxins once infected by a bacteriophage carrying tox+ gene)
○ C. ulcerans
○ C. pseudotuberculosis

CORYNEBACTERIUM SPECIES
Corynebacterium species OTHER SIGNIFICANT CHARACTERISTICS ASSOCIATED INFECTION LABORATORY DIAGNOSIS

Corynebacterium Diphtheriae
Premier pathogen of the group
Humans are the only natural hosts
Found in the URT
MOT: Spread by droplet infection or
hand-to-mouth contact
Non-motile
DIPHTHERIA TOXIN Gram (+) rods
Toxicity is due to its ability to block protein synthesis Highly pleomorphic and appear in palisades or as individual cells lying at
sharp angles to one another in V and L formations described as “Chinese
Toxin production depends on a lysogenic state in which the bacterium is Characters”
infected with a bacteriophage that carries the tox+ gene, which codes for
the production of diphtheria toxin Often stain irregularly especially when stained with Methylene Blue,
giving a beaded appearance
Only toxin-producing C. diphtheriae causes the infection diphtheria,
however, can still be converted into diphtheria toxin-producing C. Metachromatic areas of the cell that stain more intense than the other parts
diphtheriae once infected with appropriate bacteriophage. are called Babes-Ernst Granules indicating accumulation of food
reserves
Production of the toxin in vitro depends on a number of environmental
conditions:
● Alkaline pH (7.8-8.0)
● Oxygen
● Iron concentration in the medium Amount of iron needed for
optimal toxin production is less than that needed for optimal
growth The toxin is released in significant amounts only
when the available iron in the culture medium is exhausted

Occurs in two (2) forms:

● Respiratory
● Cutaneous

Incubation period: 2-5 days, characterized by low grade fever, malaise

and a mild sore throat

Most common site of infection: tonsils or pharynx

Organisms rapidly multiply on the epithelial cells and trigger an Non-spore-former or nonsporulating
inflammatory reaction
Non-motile
Infecting toxigenic strain produces toxin locally, causing tissue necrosis
and exudate formation Facultative anaerobe
 GENIO, EMMANUEL JOHN A.
BSMT L3

The combination of cell necrosis and exudate forms a very tough BIOCHEMICAL REACTIONS:
gray to white pseudomembrane that attaches to the tissues ● Ferments glucose and maltose
● May appear on the tonsils and then spread downward into the ● Reduces nitrate to nitrite
larynx and trachea

Potential for suffocation if the membrane spreads and blocks the air
passage or if dislodged

Toxin is also absorbed and produces systemic effects involving the

kidneys, heart and nervous system

Death is often a result of cardiac failure

Treated by prompt administration of antitoxin (commercial diphtheria

antitoxin is produced in horses)

Antibiotics have no effect on toxin that is already circulating, but

serve to eliminate the focus of infection as well as to prevent the spread
of the organism

SPECIAL CONSIDERATION:
Identification of an isolate as C. diphtheriae does not mean that the
patient has diphtheria

Diagnosis of diphtheria depends on showing that the isolate

produces diphtheria toxin

DRUG OF CHOICE: Penicillin

PREVENTION: DPT VAX

Tinsdale Agar
A brown halo surrounding the colony is
useful differentiating feature
C. diphtheriae can be differentiated from the
other 2 species by its lack of urease production
Three (3) biotypes of C. diphtheriae
● Mitis
● Intermedius
● Gravis
In Vivo Test
RARELY DONE
Carried out in guinea pigs
Approximately 24 hours prior to the test, one
guinea pig is injected with diphtheria antitoxin
The next day, the protected guinea pig and an
untreated guinea pig are injected with a
suspension of the suspected organism prepared
from Loeffler slants
If isolate produces diphtheria toxin, the
untreated animal will die within 3-5 days, and
the antitoxin treated animal will survive
GENIO, EMMANUEL JOHN A.
BSMT L3
CULTURAL MEDIA
Cystine-Tellurite Agar (CTA) Loeffler Serum Agar Pai Slant
Tellurite salts inhibit the growth of most normal respiratory flora Characteristic microscopic morphology -
is demonstrated well
TOXIGENICITY TESTS
Elek Test Shick’s Test
Organisms (controls and unknowns) are streaked on media of low iron content to optimize toxin production A skin test which determines
whether the patient is immune or
Organisms are each streaked in a single straight line parallel to each other and 10 mm. apart on an Elek Plate (KL not
Virulence Agar)
A filter paper strip impregnated with diphtheria antitoxin is laid along the center of the plate on a line at
right angles to the lines of control and unknown organisms
The plate is incubated at 35˚C and examined after 18, 24 and 48 hours
The white precipitin lines start about 4-5 mm. from the filter paper strip and are at an angle of about 45 degrees to
the line of growth
If isolate is (+) for toxin and is placed next to the (+) control, the toxin line of the (+) control should join the
line of the (+) unknown to form an “arch of identity”
 GENIO, EMMANUEL JOHN A.
BSMT L3

OTHER CORYNEBACTERIUM SPECIES

● C. ulcerans (subsp of C. diphtheria)

● C. pseudotuberculosis (subsp of C. diphtheria)
● C. xerosis
● C. jeikeium
● C. auris
● C. pseudodiphtheriticum
● C. urealyticum
● C. striatum
● C. kutscheri

GENUS LISTERIA
Listeria species Other significant characteristics Associated infection Virulence factors Laboratory diagnosis
Listeria monocytogenes Serotype la, lb, lVb Causative agent of LISTERIOSIS Listeriolysin O SPECIMEN
● Blood
Only human and animal pathogen among the 7 Pregnant Women Catalase ● CSF
species ● Commonly seen during 3rd trimester ● Swabs of lesions
● Responsible for spontaneous abortion Superoxide Dismutase
Has been recovered from soil, water, vegetation and and stillborn neonates MOTILITY / APPEARANCE IN CULTURE MEDIA
animal products ● Flu-like 2. Phospholipase C Gram (+) coccobacillus that often appears coccoid in
nature
Associated with the ingestion of contaminated food Newborn (granulomatosis infantisepticum) Surface Protein p60 Non spore-former
with subsequent systemic spread via the intestines ● Extremely serious: Fatality rates are high

● Two forms of neonatal listeriosis
○ Early onset ----- sepsis .
○ Late onset ----- meningitis
Immunosuppressed Host
● Meningitis
● Meningoencephalitis
● Sepsis
Infection of apparently healthy individuals may occur
via the intestinal tract when they eat contaminated
food
Outbreaks have occurred as a result of eating
contaminated cheese, coleslaw and chicken
Manifestation is nearly always meningitis and the
fatality rate is high
GENIO, EMMANUEL JOHN A.
BSMT L3
Internalin Aerobic
β-hemolytic on Sheep’s Blood Agar
Colonies are small, smooth and translucent
showing a very narrow zone of β-hemolysis
Motile at room temperature
Wet mount: exhibits “tumbling motility”
Use of motility medium demonstrates the
characteristic “umbrella” pattern when the
organism is incubated at room temperature
(25˚C) but not at 35˚C
BIOCHEMICAL TEST REACTION
CAMP Test (+) - Maybe confused with
Streptococcus agalactiae because of its striking
resemblance
● similar to S. agalactiae when S. aureus is
utilized to augment hemolysis
● More pronounced CAMP reaction is seen
with L. monocytogenes when Rhodococcus
equi is used in place of S. aureus (NOT
USUALLY DONE BECAUSE Rhodococcus is
rarely found and is expensive)
Catalase (+) which differentiates it from
Streptococcus
Hippurate hydrolysis (-)
COLD ENRICHMENT
Can grow at 4˚C, an unusual characteristic
Inoculation of specimen into broth and
incubated at 4˚C for several weeks is best to
isolate the organism from clinical specimens
(REF TEMPERATURE)

Erysipelothrix specie
Erysipelothrix rhusiopathiae
GENIO, EMMANUEL JOHN A.
BSMT L3
GENUS ERYSIPELOTHRIX
Other significant characteristics Associated infection Laboratory diagnosis
Only species in the genus Septicemia SPECIMEN
Tissue biopsy or aspirate from skin lesions
Commensal or a pathogen in a very wide variety of vertebrates Endocarditis
and invertebrates BIOCHEMICAL TEST REACTION
Erysipeloid ● H S (+) in TSI or KIA
Domestic swine are the major reservoir ● Localized skin infection that resembles ● Catalase (-)
streptococcal erysipelas ● Indole (-)
Human cases are rare, with infections resulting from ● Lesions are seen on hands and fingers: organisms ● Oxidase (-)
occupational exposure usually are inoculated through work activities
● Those whose work involves handling fish and animal ● Infected area is painful and swollen and gives rise to a CULTURAL CHARACTERISTICS/ APPEARANCE
products are most at risk characteristic lesion ● Gram (+) pleomorphic rod that has a tendency
○ A sharply defined, slightly elevated, to form long filaments
The usual route of infection is through cuts or scratches on purplish red zone that spreads peripherally ● Non motile
the skin as discoloration of the central area fades ● Non-spore former
● Non-hemolytic or alpha hemolytic
Resistant to salting, pickling and smoking
AST
Survives well in environmental sources such as water, soil and ● Neomycin Resistant
plant material ● Vancomycin Resistant