GENUS

MYCOBACTERIA
 Mycobacterium tuberculosis
(Koch bacilli, AFB, or Tubercle bacilli)
◼ AFB and gram (+) bacilli
◼ aerobic, NM, NS, NE
◼ cell wall has mycolic acid
◼ requires whole egg for growth
◼ produces “cauliflower like growth on culture
medium
◼ luxuriant growth (eugonic)
◼ causes tuberculosis in man
Culture Media
l. Non-selective
◼ Lowenstein Jensen (LJ)
◼ Petragnani
◼ American Thoracic Society
Components of these non-selective media:
◼ coagulated egg, glycerol, salt, potato, flour and
malachite green
Incubation: 37 C for 6-10 weeks, examine weekly
for growth
ll. Agar-based Media
◼ Middlebrook 7H10 & Middlebrook 7H11
Composition: glycerol, oleic acid, vitamin,
salt, catalase, biotin, cofactor, oleic
acid,and albumin
lll. Selective
◼ Gruft Modified Lowenstein Jensen
◼ Selective Middlebrook 7H11
◼ Mitchison Selective 7H11
Laboratory Tests
◼ Examination of Acid fast Stained Smear
Reporting in AFS:
1-2 AFB/slide = report number of bacilli
3-9 AFB/slide = Rare (+)
10 or more/slide = few (++)
1 or more/OIO= numerous (++++)
◼ Examination of Fluorochrome-stained Smear
◼ Test for Pigment Production
◼ Niacin (+): M. simiae, M. marinum, M. chelonei
◼ Nitrate Reduction (+): M. kansasii, M. szulgai,
M. fortuitum, M. flavescens
◼ Tween 80 differentiates M. gordonae (+) from
M.scrofulaceum (-)
◼ Catalase and Heat Stable Catalase
◼ Pyrazinamidase
◼ Arylsulfatase Test- differentiates M. fortuitum-
chelonei (+) from M. phlei (-)
◼ Iron Uptake Test M. fortuitum (+)
◼ NaCl tolerance test differentiates M. flavescens
(+)
from M. szulgai (-)
◼ PCR
Skin (Tuberculin) Tests
◼ Mantoux
◼ Von Pirquet
◼ Vollmer Patch
◼ Moro Percutaneous
◼ Tine Test
MATERIALS FOR TUBERCULIN TEST
◼ Old Tuberculin (OT)
◼ Purified Protein Derivative (PPD)
Mycobacterium leprae
(HANSEN’S BACILLUS)
◼ exhibit cigar-packet arrangement
◼ non-culturable on artificial media but can be
maintained in the footpads of armadillo
◼ it causes leprosy

Two Types of Leprosy

1. Paucibacillary (Tuberculoid leprosy)
2. Multibacillary (Lepromatous leprosy)
leprosy rash
Loss of digits due to multibacillary
leprosy
Mycobacterium bovis
◼ causes tuberculosis in cows
◼ source of Bacille Calmette Guerin (BCG)
vaccine
Mycobacterium ulcerans
◼ biochemically inert
◼ causes BURULI ulcers
◼ requires 6-12 weeks to grow at 24C and at
37 C
Mantoux Tuberculin Test
RUNYON’S CLASSIFICATION
l. PHOTOCHROMOGENS
M. szulgai -photochromogens at 25C
M. kansasii
M. simiae
M. marinum
ll.
◼ M. SCOTOCHROMOGENS
scrofulaceum
◼ M. szulgai – scotochromogens at 35-37 C
(+) to Nitrate, slow (+) for Tween 80,
can’t grow in 5% NaCl
◼ M. gordonae –tap water bacilli
(+) to Tween 80 and Heat Stable Catalase
◼ M. xenopi – slow growing organism that produces small
colonies
On Cornmeal agar growth is described as “Bird’s nest”
(-) Niacin and Nitrate, (+) Catalase , (+) in Arylsulfatase
after 2 weeks
Identified as the agent of Pulmonary Infections with Pre-
existing Lung Pathogens
◼ M. thermoresistible
◼ M. flavescens – normal flora that can grow in 5% NaCl
lll. NON-PHOTOCHROMOGENS
◼ M. malmoense (-) to Niacin and Nitrate,
(+) Tween 80 and Heat Stable Catalase
◼ M. haemophilum – requires Hemin for
growth
◼ M. avium-intracellulare – Battery bacillus
all tests are (-) except for the Heat Stable
Catalase and Pyrazinamidase Test which
are (+)
◼ M. gastri – may be isolated from gastric
lavage
lV. RAPID-GROWERS
◼ M. fortuitum (+) Nitrate and Iron Uptake
◼ M. chelonei (-) Nitrate and Iron Uptake
Note: Both can grow on McConkey agar
◼ M. smegmatis – may be confused with
M. tb in urine culture
◼ M. phlei