METHODS OF STUDYING BACTERIA

Shape and arrangement

Gram reaction

Biochemical and growth characteristics

Antigenic structure

Classification of bacteria

Aerobic and anaerobic

Spore formation

Biochemical profile

What is included in biochemical and growth characteristics?

Staphylococci

Coccus clusters

Streptococci

Coccus chains

Diplococci

Coccus pairs

Neisseriae

Coccus kidney bean-shaped, in pairs

Bacillus anthracis

Bacillus square ends

Mycobacterium tuberculosis

Bacillus rounded ends

Corynebacteria

Bacillus club-shaped

Fusobacteria

Bacillus fusiform

Vibrio

Bacillus comma-shaped

Treponema pallidum

Spirochete tightly coiled

Borrelia

Spirochete relaxed coiled

Motile

Those with flagella

Non-motile

No flagella but may show brownian motility

Brownian movement

Movement of non-motile organism due to the movement of molecules surrounding them

Staining

Process of artificially coloring the organism with dyes/stains

Simple staining

Uses only one dye and the dye is also a resulting color

Indirect/relief/negative

Background is stained, not the organism

Special staining

Use to demonstrate special features of the cell

Hiss

Anthony's

Tyler

Muir

Capsular stains

Schaeffer and fulton

Dorners

Wirtz

Conklin

Spore stains

Gray

Leifson

Fisher and conn

Flagellar stain

Albert’s

Ljubinsky

Neisser

Ponder

Methylene blue

Lindergran

Burke's technique

Metachromatic granule stain (ALNPMbLB)

Fuelgen

Nucleic acid stain

Wayson

Polar bodies stain

Levaditi

Whartin stary

fontana-tribondeau

Spirochete stain

Differential staining

Utilizes more than one dye, used to differentiate one organism from another

Gram staining

Acid fast stain

2 most common differential staining

Primary dye in gram staining

Crystal violet

Gram's iodine

Mordant stain of gram staining

95% Alcohol or acetone

Decolorizer stain

Safranin

Secondary dye for gram staining

Mycobacterium tuberculosis

Acid fast stain is used to identify what?

Gram-positive

Resists decolorization and retains purple stain

Gram negative

Can be decolorized and stains red

Colorless

Decolorizer result for gram negative

Red

Gram negative counter stain result

Neisseria

Branhamella

Moraxella

Veillonella

All cocci are gram positive except

Mycobacterium

Corynebacterium

Clostridium

Bacillus

Erysipelothrix

Lactobacillus

Listeria

Actinomyces

Nocardia

(Gardnerella)

All bacilli are gram negative except (BL2AC2MEN)

Actinomyces

Streptomyces

Yeast

Mold

Higher forms of gram positive organisms

Negative

All spiral organisms are reported as gram…

Positive

All cocci are gram…

Negative

All bacilli are gram…

Positive

Higher forms of organisms are gram…

Rickettsiae

Chlamydia

Mycoplasma

Spirochetes

Not gram stained are:

Intracellular

Why is rickettsiae not gram stained?

Intracelullar

Why is chlamydia not gram stained?

Wall less

Why is mycoplasma not gram stained?

Staphylococcus

Streptococcus

Enterococcus

Readily gram-positive cocci

Neisseria

Readily gram-negative cocci

Bacillus

Corynebacterium

Clostridium

Listeria

Actinomyces

Nocardia

Readily gram-positive rods

Escherichia

Salmonella

Readily gram-negative rods (pathogenic inside and outside intestinal tract)

Shigella

Vibrio

Camplyobacter

Helicobacter

Readily gram-negative rods (pathogenic inside intestinal tract)

Overcolorization

Use of acidic gram’s iodine

Reasons why gram (+) becomes gram (-)

Undercolorization

Thick bacterial smear

Reasons why gram (-) becomes gram (+)

Klebsiella

Enterobacter

Serratia

Pseudomonas

Proteus

Providencia

Morganella

Bacteroides

Readily gram-stained pathogenic outside intestinal tract (KESPPPMB)

Haemophilus

Legionella

Bordetella

Readily gram-stained respiratory tract organisms (HLB)

Brucella

Francisella

Pasteurella

Yersinia

Readily gram-stained organisms from animal sources (BFPY)

Acid fast stain

Organisms that are very hard to stain but once stained they are difficult to decolorize due to mycolic acid/hydroxymethoxy acid that envelopes the bacteria

Mycolic acid/hydroxymethoxy acid

Acid fast stain are organisms that are very hard to stain but once stained they are difficult to decolorize due to what?

Mycobacterium

All bacteria are non-acid fast except

Nocardia

Slightly acid fast

Tergitol

Addition of what wetting agents is used to facilitate acid fast staining?

Decolorizer

Most crucial step of gram staining

Carbolfuschin

Primary stain of AFB staining (Ziehl neelson/hot method)

Steaming process

Mordant stain of AFB staining (Ziehl neelson/hot method)

Acid alcohol

Decolorizer of AFB staining (Ziehl neelson/hot method)

Methylene blue

Counterstain of AFB staining (Ziehl neelson/hot method)

Carbolfuschin

Primary stain of AFB staining (Kinyoun’s/cold method)

Tergitol

Mordant stain of AFB staining (Kinyoun’s/cold method)

Acid alcohol

Decolorizer of AFB staining (Kinyoun’s/cold method)

Methylene blue

Counterstain of AFB staining (Kinyoun’s/cold method)

Red

Acid fast appears what against blue background?

Blue

Non-acid fast appears what against blue background?

Malachite green

May be used as a substitute to methylene blue counterstain

Red

Acid fast appears what against green background?

Blue

Acid fast appears what against green background?

Pappenheim's method

To differentiate M.smegmantis blue from M.tuberculosis red

M.smegmantis blue from M.tuberculosis red

Pappenheim's method differentiate what?

baumgarten’s method

To differentiate M.leprae red from M.tuberculosis blue

M.leprae red from M.tuberculosis blue

Baumgarten’s method is used to differentiate what?

Fite faraco’s method

For M.leprae, hematoxylin as counterstain

M.leprae

Fite faraco’s method is used for detection of what?

Hematoxylin

Fite faraco’s method uses what counterstain?

Kinyoun's method

Acid fast organisms in tissues is best stained using what?

65 degc for 2 hrs

AFB smear can be fixed using a slide warmer set at what?

Culture media

Contains the nutritional requirements for bacterial growth

Culture

Growth of microorganism in culture medium

Liquid

No solidfying agent

NB

BHI

APW

Liquid culture

0.5-1%

Semi-solid contains how much agar?

SIM

Example of semi-solid

2-3%

Liquefiable solid contains how much agar?

EMB

Example of liquefiable solid

Non-liquefiable

When heated again will no longer liquify

Rice medium

Rice grain agar

Example of non-liquifiable culture

Biphasic media

both solid and liquid

HBT

Example of biphasic media

Red algae

Agar is usually derived from what?