Exercise 4: Acid-Fast and Spore Stain

An acid-fast stain is an example of what type of stain? Why?

differential; differentiates organisms based on some characteristics that are intrinsically part of the cell

A endospore stain is an example of what type of stain?

structural; used to identify structures that are separate from the cell

The acid-fast stain is also known as:

the Ziehl-Neelsen Method

Acid-fast staining is used for what type of organisms?

those with high mycolic acid content in their cell walls

What is used to force the primary dye to penetrate the cell wall in acid-fast staining?

steam

fastness means:

to retain the dye when challenged by a decolorizer

What are the dyes/reagents used in acid-fast staining (in order)

-carbol fuchsin

-steam

-acid alcohol

-methylene blue

in acid-fast, carbol fuchsin is the:

primary dye

in acid-fast staining, which dye is "steamed?"

carbol fuchsin

in acid-fast, the acid alcohol is the:

decolorizer

in acid-fast, methylene blue is the:

secondary dye/counterstain

Acid-fast cells retain what dye and why?

retain carbol fuchsin b/c of the high wax content

Non acid-fast cells well stain what color and why?

will decolorize at acid alcohol step and will stain blue b/c it does not have high wax content

What are examples of acid-fast bacteria?

mycobacterium such as M. tuberculosis and M. leprae

M. tuberculosis is the causative agent of:

tuberculosis (TB)

M. leprae is the causative agent of:

leprosy

Spore staining is also known as:

the Schaeffer-Fulton Method

what does spore staining do?

detects dormant forms of bacteria

endospores=

dormant forms of bacteria

Can endospores be released? If so, what are they called?

yes; free spores

What is used to force the primary dye into the endospore?

steam

Stained spores are resistant to:

decolorization

What are the dyes/reagents used for spore staining (in order)

-malachite green

-steam

-distilled water

-safranin

In spore staining, the primary dye is:

malachite green

in spore staining, the decolorizer is:

distilled water

in spore staining, the secondary dye/counterstain is:

safranin

The difference between a endospore and a free spore is:

the stage of sporulation

Why are spores produced?

In response to environmental conditions

Sporulation=reproduction, T or F?

false

the endospore is the same structure as the cell it originated from, T or F?

false; it is a completely different structure from the cell

endospores stain what color?

green

free spores stain what color?

green

in spore staining, cells stain what color?

red/pink

If no spores are present, what color cells will be observed?

red/pink cells ONLY

What type of bacteria produce spores?

gram-positive ROD shaped cells

What are some examples of spore form bacteria>

-B. anthracis

-C. botulinum

-C. tetani

-C. difficile

B. anthracis is the causative agent for:

anthrax

C, botulinum is the causative agent for:

botulism

C. tetani is the causative agent for:

tetanus

C. difficile is the causative agent for:

colitis (C. diff)