Dr Mohammad Al-Tamimi, MD, Master Biomed, PhD

Classification
Gram-Positive Non-Spore-Forming
Bacilli
Medically important genera:
Listeria monocytogenes
Corynebacterium
Mycobacterium
Actinomycetes
Nocardia
1. Listeria monocytogenes
 General Characteristics
Small gram-positive rods arranged in single or short chains
Non-spore-forming, non-acid fast and don not branch
beta-hemolytic
Catalse-positive, oxidase negative
Motile: 1-4 flagella
No capsules
Facultative anaerob
Ferment glucose
Resistant to cold, heat, salt, pH extremes and bile
Multiply at refrigerator temperatures (4oC)
Virulence attributed to ability to replicate in the cytoplasm of
cells after inducing phagocytosis; avoids humoral immune system
 Epidemiology
one of the most virulent food-borne pathogens, with 20 to 30
percent of clinical infections resulting in death
Primary reservoir is soil , water; animal intestinal tract of mammals
& birds (especially chickens)
Can contaminate foods and grow during refrigeration
Transmission:

1. Dairy products, soft cheeses & unwashed raw vegetables

2. Raw or undercooked food of animal origin (luncheon meats, hot

dogs, other meat sources)
3. Transplacental to the fetus by mother blood or during birth
Often mild or subclinical in normal adults, but can induce sever
disease in immunocompromised patients, fetuses and neonates
California outbreak (29 deaths), 1985, Mexican soft cheese, due to
post-paustearization contamination
 Pathogenesis
1. Internalization into phagocytes
(internalin) by vacuole
formation
2. Secretion of listeriolysin O
(pore forming cytotoxin)
3. Polymerization of actin
filaments by Act A to form a
bacterial tail which trail the
bacteria as it moves
4. Movement and multiplication
5. Protrude to the next cell taking
the original membrane to avoid
the immune system
Clinical Presentation: Listeriosis
Neonates, elderly & immunocompromised
Granulomatosis infantiseptica
1. Transmitted to fetus transplacentally
2. Early septicemic form: 1-5 days post-partum
3. Delayed meningitic form: 10-20 days following birth
Adult septicemia
1. Gastrointestinal symptoms (nausea, vomiting, abdominal
pain, diarrhoea)
2. Septicemia induce fever and malaise
3. CNS infection leads to meninigitis and encephalitis
Laboratory Diagnosis
1. Gram stain:
Small gram-positive rods arranged in single or short
chains
Non-spore-forming, non-acid fast and don not branch
2. Culture:
Specimens: blood, CSF, biopsy
Culture of choice: 5% sheep blood, chocolate agar, or using selective
agar
Incubation conditions:
Temperature 35 C
Ambient air or 5-10% CO2
Time 24 hours
Colonial appearance: small,
white, smooth, translucent,
moist, beta-hemolytic
3. Biochemical tests:
Catalse positive
Oxidase negative
Nitrate reduction negative
Glucose fermentation positive
4. Motility test:
1. Direct wet mount: end-over and tumbling motility
when incubated in nutrient broth at room
temperature for 1-2 hours
2. Umbrella-shaped movement pattern after
overnight incubation at room temperature of a
culture stapped into semi-solid agar
5. Cold enrichment:
The specimen is placed in a nutrient broth at 4C for several weeks,
the broth is subcultured at several intervals to enhance recovery

6. Antibiotics sensitivity:
Antibiotics resistance is rare, routine sensitivity testing is not
indicated

7. Other tests: (not commonly used)

Rapid diagnostic tests using ELISA available
Antilisterolysin O IgG antibodies can be detected
The isolation of a small gram-positive catalse-
positive rods with narrow zone of beta-hemolysis
isolated from blood or CSF is a strong indicator of
listeriosis
Treatment and Prevention
Ampicillin and trimethoprim-
sulfamethoxazole
Prevention pasteurization and cooking
2. Corynebacterium Diphtheria
Introduction
Gram-positive irregular bacilli
Characteristically, they possess irregular swellings at
one end that give them the club-shaped appearance
v or k or L shape. Chinese letter pattern with angular
arrangement
Alberts stain green and bluish black
Nonmotile, noncapsulated, nonsporing and
pleomorphic
It is spread by droplets or by contact
Pathogenesis
The bacilli then grow on
mucous membranes or in
skin abrasions, and those that
are toxigenic start producing
toxin.
The bug is not invasive, but
its toxin spreads through the
body causing heart and nerve
damage.
Diphtheria toxin, is a heat-labile
AB exotoxin (Bind and Attack)
that inactivates EF-2 and thus
blocks protein synthesis.
Pathology
Diphtheria toxin is absorbed into the mucous membranes and
causes destruction of epithelium and a superficial inflammatory
response.
The regional lymph nodes in the neck
enlarge, and there may be marked edema of the entire neck,
with distortion of the airway, often referred to as bull neck
clinically.
Necrosis in heart muscle (myocarditis), liver, kidneys (tubular
necrosis), and adrenal glands, sometimes accompanied by gross
hemorrhage.
The toxin also produces nerve damage (demyelination), often
resulting in paralysis of the soft palate, eye muscles, or
extremities.
Clinical Manifestations
2 stages of disease:
1. Local infection upper respiratory tract
inflammation
sore throat, nausea, vomiting, swollen lymph nodes;
pseudomembrane formation can cause asphyxiation

2. Diptherotoxin production and toxemia

target organs primarily heart and nerves
Diagnostic Laboratory Tests
Sample collection: Throat swab or swab from membrane
Microscopy: Gram stain and Alberts stain
Culture: Selective medium such as a tellurite plate and
incubated at 37C in 5% CO2.
Eleks test for toxin detection: filter paper saturated with
antitoxin is placed on agar plate with 20% horse serum
-bacterial culture streaked at right angles to filter paper
Corynebacterium diphtheriaecoloniesontelluritebloodagar.
Prevention and Treatment
Active immunization in childhood with diphtheria
toxoid yields antitoxin levels that are generally
adequate until adulthood. (DaPT)
Rapid suppression of toxin-producing bacteria by
antimicrobial drugs
Antimicrobial drugs (penicillin,
erythromycin) inhibit the growth of
diphtheria bacilli.
Early administration of specific antitoxin
3. Actinomycetes
Introduction
Actinomycetes are a family of bacteria that form long,
branching filaments that resemble the hyphae of
fungi
Include:
Actinomyces
Norcadia
Streptomyces
Actinomyces
A. israelii most common
Strict anaerobe
Normal flora on mucous membranes and GIT
Significant opportunist pathogens
Disease: Actinomycosis
Abscesses and swelling formed at site of infection.
Diagnosis can be made upon microscopic examination of pus.
Has sulphur granules
Laboratory diagnosis:
After using ultrasound to locate an abscess, the abscess is drained
(needle aspiration) and associated sulfur granules.
This fluid is then grown (cultured) in the laboratory with absence of
oxygen (anaerobic bacterium).
 Nocardia

Nocardia infection are rare

Opportunistic: 60% of all reported nocardiosis is associated
with preexisting immune dysfunction
Lesions usually occur on the extremities, most often on the
feet .They appear as localized swollen nodules that slowly
enlarge.
Multiple abscesses form, and draining sinuses open to the
surface and discharge pus and granules.
Pulmonary: coughing blood, chest pain upon breathing
Brain: fever headache loss of neurological function
Skin: ulcers and/or nodules
Streptomyces

Streptomyces are the most widely studied

and well known genus
Inhabit soil important decomposers
Rarely pathogenic
Produce more than half of the world's
antibiotics streptomycin and erythromycin