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Cultural Characteristics
requires increase CO2 (8-12%)
requires complex media
Agar Base Media (Middle Brooke 7H-10)
Egg Base Medium (Petragnani, LJM)
growth is relatively slow (3-4 weeks
incubation)
additives are used to suppress the growth
of other microorganisms
colonies are dry, rough, granular
(Cauliflower like)
Colonies of M. tuberculosis
Lo¨wenstein-Jensen Medium
DIAGNOSIS: Epidemiology
A. Lepromen test
a skin test using sterile extract from Lepromatous Tuberculosis is endemic worldwide, less frequent in
nodule developed countries
2 types: Early (Fernandez rection, 24-48 hours) Seen from a worldwide perspective, tuberculosis is still a
Late (Mitsuda, 3-4 weeks) major medical problem.
estimated that every year approximately 15 million persons b. Neutral red dye test
contract tuberculosis and that three million die of the 4. Animal inoculation test
disease. 5. Tuberculin test
the main source of infection is the human carrier. a. OT (old original tuberculin)
transmission of the disease is generally direct, in most cases b. PPD (Purified Protein Derivative)
by droplet infection. positive test: past infection, close association
Indirect transmission via dust or milk (udder tuberculosis in with infected patient
cattle) is the exception rather than the rule.
incubation period is four to 12 weeks. Methods (tuberculin skin test)
PREVENTION
prophylactic BCG
avoid contact with infected person
Isolation and treatment of cases
LABORATORY DIAGNOSIS
Specimens: Sputum (early morning sputum) (inoculated in
Middle Brock 7H) biopsy material, pleural fluid, synovial fluid,
gastric content
1. Direct smear - AFB staining
2. Concentration technique (use of digesting
substance: 4% NaOH, 6% oxalic acid, 4% sulfuric
acid)
3. Virulence test
a. Serpentine cord formation
CORYNEBACTERIA GROUP
Gram-positive, aerobic, non-motile, non-sporeforming,
uncapsulated bacilli
pleomorphic, club-shaped
“Babes-Ernst” metachromatic granules (beaded
appearance)
arrangement: X, V and L, palisade, Chinese letter or
character
DIAGNOSIS:
1. Stained smear
a. (Grams, Methylene Blue)
2. Culture/Cultivation
Loeffler medium
BAP
Corynebacterium diptheriae PAIS coagulated Egg medium
Kleb’s Loeflers bacillus Loefler’s serum slant (enhance
causative agent of Deptheria pleomorphism)
produces 3 varieties in Tellurite Agar Tellurite Agar (best for colonial morphology)
1. C. deptheriae var. gravis a. Cystine Tellurite Blood Agar
2. C. deptheriae var. mitis b. McLeods Potassium Tellurite Agar
3. C. deptheriae var. intermedius
*mode of transmission Serological test/Toxigenicity
droplet nuclei from infected patient a. Schick’s test (Susceptibility test)
contact from coetaneous foci of infection 1 arm injected with 0.1 ml unheated toxin
the other arm with heated toxoid
Pathogenesis indicated by redness and swelling
Deptheria is only caused by C. diptheriae infected Interpretations
with bacteriophage carrying toxigene (Lysogenic (+) no antibody, susceptible to infection
strains) (-) has antibody, protected from infection
due to the powerful exotoxin (more poisonous than b. Elek test (neutralization/precipitation reaction)
that of cobra venum) c. Elek-Ouchterlony immunodiffusion test
Extracellular toxin consists of two functionally distinct d. Animal inoculation
fragments, A (toxic activity) irreversibly blocks
protein synthesis causing cell death and B (binding PREVENTION:
to receptors of target cells) active and passive immunization
supplemental antibiotics (eliminate the carrier state of
Pseudomembrane formation toxigenic strains during epidemics)
due to production of serum exudate and cellular
infiltrate of mucous membrane in the pharynx Other species
(causes respiratory obstruction 1. Deptheroids
Diptheria-like organisms
non-pathogenic, found in the throat
2. Corynebacterium minutissimum b. Pulmonary anthrax (5%)
causative agent of superficial infection of the also known as “Wool’s sorters disease”
auxiliary and pubic skin caused by inhalation of spores
infected skin when exposed to UV produces c. Intestinal anthrax (rare)
pink fluorescence obtained from ingestion of improperly
cooked infected meat (affect 6 hours after
ingestion)
DIAGNOSIS:
Specimens: blood, sputum, swab from pustules
1. Stained smear (Grams,
Immunofluorescence)
2. Culture (no special medium)
3. Animal inoculation
4. String of Pearl test
5. Ascoli test
precipitation test
ascoli is from the extract of infected
tissues
Other species
1. Bacillus thuringensis
used in biological control of mosquitoes
source of Bt toxin for GMO’s
2. Bacillus cereus
motile, non-capsulated, Beta-hemolytic
can cause food poisoning (e.g. fried rice)
two forms of infection
a) Emetic form
1. Bacillus anthracis o manifested by nausea, vomiting.
important species, highly pathogenic Abdominal cramp (recovery: 24 hours,
large, encapsulated, none-motile Gram (+) bacilli self- limiting)
known as bamboo fishing rod appearance with o incubation period: 1 -5 hours
centrally located spores o associated with fried rice
disease of animals, humans are infected incidentally b) Diarrheal type
Pathogenicity o associated with meat dishes and
causative agent of anthracis/anthrax sauces
incubation period: 6 weeks o profuse diarrhoea with abdominal
spread rapidly via lymphatics, then to blood cramps
pathogenicity is due to presence of capsule and o incubation period: 1 – 24 hours
toxin production o also associated with eye infection:
three proteins of Anthrax toxin Severe Keratitis
1. PA (protective antigen)] Endophthalmitis
2. EF (Edema factor) 3. Bacillus steathermophilus
3. LF (lethal factor) current name: Geobacillus steathermophilus
Clinical type of anthrax used to determine the efficacy of autoclaving and
a. Cutaneous anthrax (95%) other sterilization procedures
malignant pustule 4. Bacillus subtilis
caused by skin contact with infected animals also known as Hay bacillus, common
may progress to systemic infection environmental contaminant
CLOSTRIDIA GROUP Clostridium tetani
most species are motile with peritrichous flagella with terminally located spore, motile
hemolytic and non-capsulated drumstick or tennis rocket appearance
found in soil, feces of horses and other animals Common source of infection: dirty pointed objects,
injured area with spores deposited on it, then
absorbed by the blood stream
Pathogenicity
produces toxins
a. Tetanus lysin (Hemolysin)
o responsible for the Beta-hemolytic
activity
DIAGNOSIS:
1. Stained smear LACTOBACILLUS SPECIES
2. Culture of contaminated food giant rod-shape bacillus (Doderlein’s bacillus)
3. Serological test normal flora of the vagina (L. acidophilus)
also inhabits the mouth and GIT (colon)
PREVENTION: produces large amount of lactic acid from CHO
boil food for 15 minutes/ heat at least 80 fermentation when immune system is suppressed,
C causes meningitis and endocarditis
DIAGNOSIS:
involves culturing the pathogen from patient stool -
detection of the cytotoxin in bacteria-free stool
filtrates on the basis of a cytopathic effect (CPE)
observed in cell cultures
Gardnerella vaginalis
a Gram-variable, nonmotile, nonencapsulated rod
bacterium.
natural habitat: vagina of sexually mature
women
pathology: vulvovaginitis (vaginosis), found in over
90% of women showing the symptoms of this
infection