Lesson 15:

Mycobacteria
(acid fast
bacilli)
 LESSON TOPIC:
MYCOBACTERIA (ACID FAST
15 BACILLI)
Learning Objectives:
At the end of the lesson, students should be able to:
1) Explain the general characteristics of the genus
Mycobacterium;
2) Identify the infections and diseases associated with acid
fast gram positive bacilli; and
3) Differentiate each specie of Mycobacteria fom each
other.
MYCOBACTERIA (ACID
FAST BACILLI)
General Characteristics of Mycobacteria
o This species of this genus are non-motile, non-spore-forming and non-
encapsulated
o They are strictly aerobic and catalase-positive, and produces Much’s
granules
o Most species are associated with diseases that develop slowly (requires
two -6 weeks of incubation) while the rapidly growing mycobacteria
species multiply in 2-3 days
o Microscopy: Slender, slightly curved or straight rods that have a
tendency to “clump”
o There are two groups: Mycobacterium tuberculosis complex (MTC)
and non-tuberculous mycobacteria (NTM)

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 MYCOBACTERIA (ACID
FAST BACILLI)
Mycobacterium tuberculosis complex (MTC)
1. Mycobacterium tuberculosis (Koch’s bacillus)
✗ Most common cause of tuberculosis
✗ Culture: Colonies are slow growing; are raised and dry and have a cauliflower-like appearance
✗ Highly resistant to drying; Droplets of dried sputum in the air may be infectious for 8-10 days
2. Mycobacterium bovis
✗ It produces tuberculosis in humans and animals (cattle, dogs, cats and swine)
✗ Its attenuated strain is used for vaccination (Bacillus-Calmette-Guerin or BCG vaccine) of
newborns against tuberculosis
✗ Acquired through ingestion of contaminated milk from infected cows or exposure to infected
animals and their carcasses

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 MYCOBACTERIA (ACID
FAST BACILLI)
Mycobacterium tuberculosis complex (MTC)
3. Mycobacterium africanum
 Associated with human cases of tuberculosis in tropical Africa

4. Mycobacterium canetti
 Grows more rapidly than M. tuberculosis (six days on solid media)
 Isolated from an AIDS patient with mesenteric tuberculosis

5. Mycobacterium microti
 Has been isolated from TB patients in both immunocompetent and immunocompromised
individuals

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 MYCOBACTERIA (ACID
FAST BACILLI)
Related Infections and Diseases
1. Tuberculosis
 A disease of the respiratory tract
 Chronic granulomatous infection which is transmitted through the inhalation of infected droplets
by means of coughing, sneezing or speaking
 Signs and symptoms: low grade fever, night sweats, fatigue, anorexia and weight loss
 Clinical diagnosis of primary tuberculosis: tuberculin test using purified protein derivative (PPD)-
injected intradermally (+ redness after 48 hours)

2. Pott’s disease
 Also known as tuberculosis spondylitis or skeletal tuberculosis of the spine
 Serious form of tuberculosis that is caused by the invasion of M. tuberculosis into spinal vertebrae

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MYCOBACTERIA (ACID
FAST BACILLI)
Non-tuberculous mycobacteria (NTM)
 Also known as the atypical mycobacteria or mycobacteria other than tuberculosis (MOTT)
 Found in the environment (soil, dust and water) and they sometimes colonize the skin, respiratory tracts
and GI tracts of healthy individuals
 Chronic pulmonary disease resembling tuberculosis is the usual clinical presentation associated with
these organisms
 Infections that are caused by these organisms are not considered as transmissible from person to person
 Incidents of acquired immunodeficiency syndrome (AIDS) have contributed greatly to the incidence and
awareness of NTM diseases

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MYCOBACTERIA (ACID
FAST BACILLI)
NTM Slow growers

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 MYCOBACTERIA (ACID
FAST BACILLI)
NTM Slow growers
1. Mycobacterium avium complex
✗ Most common cause of pulmonary infections in humans that is similar to tuberculosis
✗ Pathogen in immunocompetent and immunocompromised (AIDS) patients
✗ It can be isolated from sputum, blood and bone marrow aspirates
✗ Species: M. avium, M. intracellulare, M. avium subsp. Paratuberculosis and M. avium subsp.
Silvaticum

2. Mycobacterium kansasii (Yellow bacillus)

✗ Second to M. avium complex as the cause of NTM lung diseases (chronic cavitary pulmonary
lesions) in humans
✗ Not normally contagious from person to person; Photochromogenic colonies

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 MYCOBACTERIA (ACID
FAST BACILLI)
NTM Slow growers
2. Mycobacterium marinum
✗ Causes diseases on fishes and can be isolated from aquariums
✗ Causative agent of “swimming pool granuloma” which is red or bluish-red
nodule on the elbow, knee, toe or finger that occur when an open wound comes
into contact with contaminated chlorinated fresh water or salt water

3. Mycobacterium ulcerans
✗ Third most common Mycobacterium species after M. tuberculosis and M.
leprae
✗ Rare cause of Buruli ulcer which is a painless nodule under the skin after a
previous trauma

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 MYCOBACTERIA (ACID
FAST BACILLI)
NTM Slow growers
4. Mycobacterium gordonae (Tap water bacillus)
✗ Contaminates tap water that is used by patients in rinsing their mouths prior to sputum collection
✗ Contaminant in the preparation of bacteriologic smears
✗ It rarely causes infection in humans

5. Mycobacterium xenopi
✗ Recovered from hot and cold water taps, especially from water storage tanks of hospitals
✗ Potential pathogen of pulmonary infection in adults
✗ Can be either non-photochromogenic or scotochromogenic

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 MYCOBACTERIA (ACID
FAST BACILLI)
NTM Rapid growers
 Considered as potentially pathogenic
 Found in dusts, soil and water system
 Enter their hosts through breaks in the skin and subcutaneous tissues by trauma, injections, surgery or
animal contact

1. Mycobacterium fortuitum
✗ Most common, rapidly growing mycobacteria that are associated with localized cutaneous and soft
tissue infections

2. Mycobacterium chelonae
✗ Mostly associated with cutaneous infections in immunocompromised persons
✗ Exhibits greater resistance to antimicrobial agents than M. fortuitum

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 MYCOBACTERIA (ACID
FAST BACILLI)
NTM Rapid growers
1. Mycobacterium abcessus subsp. abcessus
✗ Reservoir: Tap water
✗ Related infection and disease: Chronic lung disease and otitis media

2. Mycobacterium smegmatis
✗ Related infections: pulmonary skin and bone infections

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 MYCOBACTERIA (ACID
FAST BACILLI)
Laboratory Diagnosis
✗ Preferred specimen:
✗ Sputum (early morning sputum 2-3 samples collected for consecutive days)
✗ Required volume: 5-10 mL
✗ Microscopy prior to culture: <10 epithelial cells and > 25 NPN (pus cells)
✗ For NTM
✗ Bronchoscopy specimens
✗ Bronchoalveolar lavage, bronchial washings and transbronchial biopsies
✗ Other specimen:
✗ Urine, fecal specimen, pleural, pericardial, peritoneal and CSF, blood, wound and tissue aspirates

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 MYCOBACTERIA (ACID
FAST BACILLI)
Laboratory Diagnosis
✗ STAINING
✗ 5,000-10,000 organisms/mL is needed to obtain a positive AFB staining
✗ Acid fastness is affected by age, culture media and exposure to UV light
✗ AFB Staining methods
✗ Kinyoun/ Ziehl-Neelsen
✗ Auramine-rhodamine fluorochrome staining (Bright yellow-orange bacilli under the fluorescent
microscope)
✗ Classification of AFB after Ziehl-Neelsen/ Kinyoun staining
✗ Distinct AFB: Mycobacterium
✗ Partially AFB: Nocardia, Gordonia, Rhodococcus, Tsukamurella, and Legionella micdadei

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MYCOBACTERIA (ACID
FAST BACILLI)
Laboratory Diagnosis

16
 MYCOBACTERIA (ACID
FAST BACILLI)
Non-cultivable NTM
1. Mycobacterium leprae (Hansen’s bacillus)
✗ It is not cultivated on any synthetic mycobacterial media
✗ Invades peripheral nerves and skin cells and becomes an obligately intracellular parasite
✗ Microscopy: Rod-shaped and exhibits a “cigar pocket” or “picket fence” arrangement
✗ Culture: Colonies exhibit growth in living tissues of the footpads of mice and armadillos
✗ Skin test: Fernandez and Mitsuda reaction

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MYCOBACTERIA (ACID
FAST BACILLI)
Non-cultivable NTM

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 MYCOBACTERIA (ACID
FAST BACILLI)
Non-cultivable NTM
1. Leprosy/ Hansen’s disease
✗ Chronic disease of the skin, mucous membranes and nerve tissues
✗ Not considered a highly contagious disease
✗ Modes of transmission: Person to person contact through inhalation (nasal
secretions), contact with infected skin, ingestion of breast milk and
transplacental transmission for infants
✗ Forms of leprosy: Tuberculoid and lepromatous

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 Thanks
!
References:
✗ Mahon, C.R., Lehman, D.C., Manuselis,
G. (2014). Textbook of
Diagnostic Microbiology (5th ed.). New
York: Saunders
✗ Bailey, W. R., Scott, E. G., Finegold, S.
M., & Baron, E. J. (1986). Bailey and
Scott's Diagnostic microbiology. St.
Louis: Mosby.
✗ Rodriguez, M.T. (2018). Review
Handbook in Diagnostic Bacteriology.
C&E Publishing Inc.
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