12 ) Mycobacterium ulcerans- (inert bacillus) inert because in most conventional test ay negative sya.

This can be found in

stagnant and tropical waters. Can cause shallow ulcer, referred to as buruli ulcer. Nonpigmented.In
Enterobacteriaciae ay meron ding inert bacillus (Shigella)
13 ) Mycobacterium xenopi- bird’s nest appearance

-rapid growing spp.:

1 ) Mycobacterium chelonae or Mycobacterium abscessus group
2 ) Mycobacterium fortuitum group
3 ) Mycobacterium smegmatis group- contains two spp.: Mycobacterium smegmatis and Mycobacterium goodii
4 ) Mycobacterium leprae- causative agent for hansen disease(leprosy)(ketong) skin lesions or nodules.
-it exist in two forms:
=tuberculoid leprosy- localized form of disease, usually merong mga lesions
=lepromatous leprosy

Isolation and identification of Mycobacteria

-specimen collection: madami hindi lang sputum because the bacteria can disseminate in other areas of the body. For the
sputum, it must be deep cough collection,early morning for 3 consecutive days. Can also submit urine, must be
collected early in the morning for 3 consecutive days
-specimen submitted can be digest and contaminate, used in specimens that have mucous for example, sputum. Example
of specimens that requires digestion and decontamination are BAL(bronchoalveolar lavage). yung ibang specimen
such as urine and tissues hindi naman mucoid so kailangan lang nila ay decontamination para mamatay ung ibang non-
Mycobacterial organisms. Meron ding mga specimen na hindi na kailangan I-digest and decontaminate, mga
specimen na galing sa normally steriled sites like blood, CSF, sinovial fluids.

Example of digestion and decontamination agents:

1 ) sodium hydroxide- concentration of 2,3,4%
2 ) N-acetyl-L-cysteine
3 ) benzalkonium chloride- combined with trisodium phosphate (TSP)
4 ) oxalic acid

Staining for acid-fast bacilli

-ziehl-neelsen method(hot) and kinyoun method(cold)- the difference is the use of mordant: mordant(hot-heat) (cold-
tergitol) , decolorizing agent
-in acid-fast staining, sometimes we use qualitative interpretation/report

Culture media:
-can be classified as egg based media and agar-based
-Egg-based media:
=lowenstein-jensen
= petragnani
=american thoracic society
-agar-based
=middlebrook 7H10
=middlebrook 7H11

Preliminary identification of Mycobacteria

-photoreactivity:
1 ) photochromogens- pale yellow-orange
2 ) scotochromogens- produce pigment in the light or in the dark(orange-red)
3 ) nonchromogenic/ nonphotochromogenic- buff to tan color

Biochemical identification
1 ) niacin accumulation- yellow colored solution
2 ) nitrate reduction- pink-red color ( added with HCl etc.)
3 ) catalase test- effervescence or formation of bubbles…other spp negative in catalase test(Mycobacterium tuberculosis):
=M. gastri
=M. haemophilum
=M. marinum