Corynebacterium

Corynebacterium diphtheria
LSS- Corynebacterium diphtheriae , also called as Klebs-Loeffler bacillus, is a gram-positive, non-
encapsulated, non-sporulated, non-motile facultative anaerobe.
- The current formulation of LSS is an incorporation of previous developed to elicit the
formation of metachromatic granules which are characteristics of c. diphtheria
- Metachromatic granules(Babes-Ernst granules) or polyphosphate- Among the functions
of poly P in cellular metabolism is its vital role in stress response and stationary-phase
adaptation, stains.
stain bluish-purple with methylene blue. The cell wall sugars include arabinose,
galactose, and mannose.
Morphology and Identification Corynebacteria are club-shape, gram positive, non spore forming
bacteria, found in chines characters-like arrangement of cells. Aerobic and non motile.

- Corynebacterium diphtheriae is a Gram-positive, rod-shaped bacterium that typically

shows positive results for nitrate reduction in laboratory tests. Nitrate reduction refers
to the ability of a bacterium to reduce nitrate (NO3-) to nitrite (NO2-) or other
nitrogenous compounds.

The positive nitrate reduction result in C. diphtheriae is due to the presence of enzymes,
such as nitrate reductase, that are capable of catalyzing the reduction of nitrate. These
enzymes are involved in the anaerobic respiration of the bacterium, allowing it to utilize nitrate
as a terminal electron acceptor in the absence of oxygen.

In laboratory tests, the nitrate reduction reaction is often detected using a nitrate test medium
containing nitrate and a reagent (such as sulfanilic acid and dimethyl-alpha-naphthylamine)
that forms a red color when nitrite is present. If the medium turns red after incubation with the
bacteria, it indicates the presence of nitrite, which suggests that the bacteria have reduced
nitrate.

- The lack of urease production in C. diphtheriae is one of the factors that help
differentiate it from other bacteria in the laboratory.
- Corynebacterium diphtheriae is catalase-positive due to the presence of the enzyme
catalase. The gene encoding catalase in C. diphtheriae is part of its genome, and the
bacterium synthesizes this enzyme as part of its metabolic processes.
- Catalase is an important enzyme for C. diphtheriae as it helps the bacterium neutralize
toxic hydrogen peroxide (H2O2), which can be produced during aerobic metabolism or
as a host defense mechanism. By breaking down hydrogen peroxide into water and
oxygen, catalase protects C. diphtheriae from the damaging effects of this reactive
oxygen species.
 - The presence of catalase in C. diphtheriae is a characteristic feature that can be used in
laboratory tests to distinguish it from other bacteria and aid in its identification.
- Glucose and maltose are both carbohydrates that can be used as energy sources by C.
diphtheriae. The bacterium possesses the necessary enzymes to metabolize these
sugars, converting them into products such as organic acids (e.g., lactic acid) and gases
(e.g., carbon dioxide). The specific fermentation pathways used by C. diphtheriae for
glucose and maltose may vary, but ultimately, these sugars are utilized to support the
bacterium's growth and metabolism.
- The ability of C. diphtheriae to ferment glucose and maltose can be detected in the
laboratory using fermentation tests. These tests typically involve inoculating the
bacterium into a medium containing the sugar of interest and observing for the
production of acid and gas as indicators of fermentation. The ability to ferment glucose
and maltose is a characteristic feature of C. diphtheriae that can help differentiate it
from other bacteria

Pathogenesis

1. C. diphtheriae is an aerobic, gram-positive bacillus. Toxin production (toxigenicity)

occurs only when the bacillus is itself infected (lysogenized) by specific viruses
(corynebacteriophages) carrying the genetic information for the toxin (tox gene).
Diphtheria toxin causes the local and systemic manifestations of diphtheria
2. What is the nasopharynx? Your nasopharynx is the top part of your throat (pharynx). It's
a muscular, box-shaped passageway behind your nose, just above the roof of your
mouth. Your nasopharynx allows air to pass from your nose into your windpipe and
eventually into your lungs
3. The disease Diphtheria is caused by C. diphtheria (toxin producer). It is a droplet
infection in which the organisms pass through the nasopharynx. Non toxic form of
Corynebacteria found in the normal microbiota are called (Diphtheroids).
HOW?
- Diphtheria-causing bacteria may produce a toxin. This toxin damages tissue in the
immediate area of infection — usually, the nose and throat. At that site, the infection
produces a tough, gray membrane made up of dead cells, bacteria and other
substances. This membrane can obstruct breathing.
- Corynebacterium diphtheriae is the bacterium responsible for causing diphtheria, a
potentially serious infectious disease. Here's an overview of its pathophysiology:
-
- 1. **Transmission**: C. diphtheriae is spread from person to person through respiratory
droplets or close contact with an infected person.
-
- 2. **Entry and colonization**: After entering the body, usually through the respiratory
tract, C. diphtheriae attaches to the mucous membranes of the throat and upper
respiratory tract.
-
- 3. **Toxin production**: C. diphtheriae produces a potent exotoxin called diphtheria
toxin. This toxin is encoded by a bacteriophage (a virus that infects bacteria) carrying the
tox gene. The toxin is released by the bacteria and can cause damage to nearby tissues.
-
- 4. **Toxin effects**: The diphtheria toxin affects protein synthesis in the host cells,
leading to cell death and tissue damage. It primarily affects the respiratory tract, causing
a characteristic pseudomembrane to form in the throat, which can obstruct the airway.
-
- 5. **Systemic effects**: The toxin can also enter the bloodstream and affect other
organs, leading to systemic complications such as myocarditis (inflammation of the
heart muscle) and neuritis (inflammation of nerves).
-
- 6. **Immune response**: In response to the infection, the body mounts an immune
response, which can lead to inflammation and further tissue damage.
-
- 7. **Complications**: Without prompt treatment, diphtheria can lead to serious
complications, including airway obstruction, myocarditis, and nerve damage. In severe
cases, it can be fatal.
-
 - Treatment of diphtheria involves administering antitoxin to neutralize the toxin, along
with antibiotics to kill the bacteria. Vaccination with the diphtheria vaccine is the most
effective way to prevent diphtheria.
The palisade (meaning "stake" in Latin) is a layer of closely packed cells found under the
epidermis on the upper layer of a leaf.

Stained Corynebacterium cells. The "barred" appearance is due to the presence of

polyphosphate inclusions called metachromatic granules. Note also the characteristic "Chinese-
letter" arrangement of cells.
Cystine-tellurite blood agar (CTBA) is a selective and differential medium used for the isolation
and identification of Corynebacterium diphtheriae, the bacterium that causes diphtheria. Here's
how CTBA is used for the detection of C. diphtheriae:

1. **Selective medium**: CTBA contains ingredients that inhibit the growth of many bacteria
while allowing for the growth of C. diphtheriae. The tellurite in the medium is toxic to most
bacteria but is reduced by C. diphtheriae, leading to the formation of tellurite crystals that
appear black or gray on the medium.
2. **Differential medium**: CTBA also contains nutrients like blood (usually sheep or horse
blood) and cystine, which support the growth of C. diphtheriae. The medium can differentiate
C. diphtheriae based on its ability to reduce tellurite and form characteristic black colonies.
3. **Detection**: When a sample containing C. diphtheriae is streaked onto CTBA and
incubated, C. diphtheriae colonies appear as black or gray colonies due to tellurite reduction.
Other bacteria that do not reduce tellurite will not form black colonies.

CTBA is a valuable tool in the laboratory diagnosis of diphtheria, as it allows for the selective
growth and identification of C. diphtheriae based on its characteristic colony morphology.
Modified Tinsdale agar is another selective and differential medium used for the isolation and
identification of Corynebacterium diphtheriae, particularly the toxigenic strains that produce
diphtheria toxin. Here's how it is used for the detection of C. diphtheriae:

1. **Selective medium**: Modified Tinsdale agar contains potassium tellurite, which is

inhibitory to most bacteria but allows for the growth of C. diphtheriae. The tellurite is reduced
by C. diphtheriae, resulting in the formation of black or gray colonies due to the precipitation of
tellurium.
2. **Differential medium**: In addition to tellurite, Modified Tinsdale agar contains other
ingredients that allow for the differentiation of C. diphtheriae based on its ability to produce
hydrogen sulfide (H2S) and form brown or black colonies.
3. **Detection**: When a sample containing C. diphtheriae is streaked onto Modified Tinsdale
agar and incubated, C. diphtheriae colonies appear as black or gray colonies with a brown halo
due to tellurite reduction and hydrogen sulfide production. Non-toxigenic strains of C.
diphtheriae may still grow but will not produce the characteristic brown halo.

Modified Tinsdale agar is useful for the selective isolation of C. diphtheriae and for the
differentiation of toxigenic and non-toxigenic strains based on colony morphology. The
presence of toxigenic strains is important for the diagnosis and management of diphtheria
cases.
Cystine is a sulfur-containing amino acid that serves as a nutrient source for many bacteria,
including Corynebacterium diphtheriae. In the context of Cystine-tellurite blood agar (CTBA),
cystine is added to the medium to enhance the growth of C. diphtheriae. Here's how cystine
enhances the growth of C. diphtheriae:

1. **Nutrient source**: Cystine provides a source of sulfur and carbon for C. diphtheriae, which
are essential nutrients for bacterial growth and metabolism. Bacteria like C. diphtheriae can
utilize cystine as a building block for proteins and other cellular components.
2. **Energy production**: Cystine can be metabolized by C. diphtheriae to produce energy
through various metabolic pathways. The sulfur in cystine can be used by the bacterium in
sulfur-containing metabolic reactions.
3. **Growth promotion**: The presence of cystine in the medium promotes the growth of C.
diphtheriae by providing essential nutrients that support bacterial replication and proliferation.
This can lead to more robust growth and the formation of visible colonies on the agar medium.
Overall, cystine enhances the growth of C. diphtheriae by providing essential nutrients and
energy sources that support bacterial metabolism and replication. Its inclusion in CTBA helps to
create a medium that is conducive to the selective growth of C. diphtheriae for laboratory
diagnosis.

Corynebacterium jeikeium is a bacterium that is part of the normal flora of human skin and
mucous membranes. However, it can also act as an opportunistic pathogen, particularly in
immunocompromised individuals or those with underlying health conditions. Here are some
key aspects of the pathogenesis of Corynebacterium jeikeium:
1. **Colonization**: Like other members of the Corynebacterium genus, C. jeikeium can
colonize the skin and mucous membranes of humans. It is often found in the axillae, groin, and
perineum, as well as on the hands and feet.

2. **Virulence factors**: C. jeikeium possesses several virulence factors that contribute to its
pathogenicity. These include surface adhesins that allow the bacterium to adhere to host
tissues, as well as enzymes that help it evade the host immune response.

Susceptibility testing for Corynebacterium jeikeium is typically performed using standard

methods such as the disk diffusion method or the broth microdilution method. Here is an
overview of how susceptibility testing is performed for C. jeikeium:

1. **Isolation of the organism**: Before susceptibility testing can be performed, the organism
must be isolated from the clinical specimen and identified as C. jeikeium using appropriate
biochemical and/or molecular methods.

2. **Preparation of the inoculum**: A standardized inoculum of the bacterium is prepared,

usually equivalent to a 0.5 McFarland standard, to ensure consistency in the test results.

3. **Selection of antibiotics**: Antibiotics commonly used for susceptibility testing of C.

jeikeium include beta-lactams (such as penicillin and cephalosporins), glycopeptides (such as
vancomycin), and macrolides (such as erythromycin). The selection of antibiotics may vary
based on local resistance patterns and the clinical scenario.

4. **Disk diffusion method**: In the disk diffusion method, paper disks containing specific
concentrations of antibiotics are placed on an agar plate inoculated with the organism. The
plates are then incubated, and the diameter of the zone of inhibition around each disk is
measured and interpreted according to established guidelines.

5. **Broth microdilution method**: The broth microdilution method involves preparing a series
of twofold dilutions of antibiotics in a microtiter plate. Each well of the plate is inoculated with
the standardized bacterial suspension, and the plates are incubated. The minimum inhibitory
concentration (MIC) of each antibiotic is determined as the lowest concentration that inhibits
visible growth.

6. **Interpretation of results**: The results of susceptibility testing are interpreted based on

established criteria for each antibiotic. The organism is classified as susceptible, intermediate,
or resistant to each antibiotic tested.

7. **Reporting**: The results of susceptibility testing are reported to the healthcare provider,
who can then use this information to guide antibiotic therapy.

It's important to note that susceptibility testing should be performed in a clinical microbiology
laboratory following standard protocols and guidelines to ensure accurate and reliable results.

3. **Infection**: In immunocompromised individuals or those with compromised skin barriers

(such as burns or surgical wounds), C. jeikeium can cause infections. These infections can range
from superficial skin infections, such as erythrasma, to more serious infections, such as
bloodstream infections (bacteremia) or infections of implanted medical devices.

4. **Antibiotic resistance**: C. jeikeium is known for its high level of resistance to many
antibiotics, including beta-lactams and macrolides. This can make infections caused by this
bacterium difficult to treat, especially in healthcare settings where antibiotic-resistant strains
are more common.

5. **Nosocomial infections**: Due to its resistance to antibiotics and its ability to survive in the
environment, C. jeikeium can cause nosocomial (hospital-acquired) infections. These infections
are often associated with the use of indwelling medical devices, such as catheters or prosthetic
joints.

Overall, while Corynebacterium jeikeium is a normal part of the human microbiota, it can cause
opportunistic infections, particularly in immunocompromised individuals or those with
underlying health conditions. Its ability to adhere to host tissues, evade the immune response,
and resist antibiotics contributes to its pathogenicity.
Vancomycin is used as a treatment for Corynebacterium jeikeium infections primarily because
of its effectiveness against Gram-positive bacteria, including multidrug-resistant strains. Here
are some key reasons why vancomycin is used for treating Corynebacterium jeikeium
infections:

1. **Broad-spectrum activity**: Vancomycin has a broad spectrum of activity against Gram-

positive bacteria, including Corynebacterium jeikeium. It is effective against many strains of
bacteria that are resistant to other antibiotics, such as methicillin-resistant Staphylococcus
aureus (MRSA) and vancomycin-resistant enterococci (VRE).

2. **Mechanism of action**: Vancomycin works by inhibiting cell wall synthesis in bacteria. It

binds to the D-alanyl-D-alanine portion of the cell wall precursor, preventing its incorporation
into the growing cell wall. This ultimately leads to cell death.

3. **Intracellular penetration**: Vancomycin has good penetration into tissues, including skin
and soft tissues, where Corynebacterium jeikeium infections commonly occur. This allows for
effective treatment of localized infections.

4. **Minimal resistance**: While some strains of bacteria, including Corynebacterium jeikeium,

can develop resistance to vancomycin, it is generally less common compared to other
antibiotics. This makes vancomycin a reliable option for treating infections caused by multidrug-
resistant bacteria.

5. **Clinical experience**: Vancomycin has been used for many years in clinical practice and
has a well-established safety profile. It is available in various formulations, including
intravenous and oral, making it suitable for different types of infections and patient
populations.

Overall, vancomycin is a valuable antibiotic for the treatment of Corynebacterium jeikeium

infections, particularly when other antibiotics are ineffective due to resistance. However, as
with any antibiotic, careful monitoring and appropriate use are essential to minimize the risk of
resistance and adverse effects.