1.

MOTILITY
A. B. C. D. E. F. HANGING-DROP METHOD USE OF SEMI-SOLID MEDIA STAINING OF FLAGELLA SEROLOGICAL TEST FLUORESCENT TEST SWARMING PHENOMENA

2. MORPHOLOGY AND STAINING 3 GENERAL WAYS A. WET MOUNT B. HANGING DROP C. FIXED STAINING

1.

CLASSIFICATION ACCORDING TO SHAPE:

A. BACILLI : ROD SHAPED B. COCCI : ROUND OR SPHERICAL C. SPIRILLA : SMALL, COMMA-SHAPED OR SPIRAL ORGANISM
VIBRIO SPIRILLUM SPIROCHETE

2. CLASSIFICATION ACCORDING TO ARRANGEMENT:

A. STAPHYLO B. STREPTO C. SARCINAE D. TETRAD E. DIPLO F. PALLISADE

PROCESS OF ARTIFICIAL COLORING OF MICROORGANISM WITH DYES OR REAGENTS IN ORDER TO FACILITATE MORPHOLOGICAL IDENTITY AND ARRANGEMENT OF BACTERIA UNDER THE MICROSCOPE.

1.

SIMPLE STAIN
-

ONE PARTICULAR STAIN OR DYE RESULTING INTO ONE COLOR ALSO KNOWN AS DIRECT STAINING AN AQUEOUS OR ALCOHOLIC SOLUTION OF A SINGLE DYE.

EX: LOFFLERS METHYLENE BLUE GENTIAN VIOLET CARBOL FUCHSIN SAFRANIN

2. DIFFERENTIAL STAIN
TWO OR MORE DYES OR STAINS ARE USUALLY EMPLOYED COMPOSITION : 1. PRIMARY STAIN 2. MORDANT 3. DECOLORIZER 4. SECONDARY OR COUNTER STAIN EX. : GRAMS STAIN, ACID FAST STAIN

REAGENTS :
1.
2. 3. 4.

CRYSTAL VIOLET GRAMS IODINE ALCOHOL AND /OR ACETONE SAFRANIN

THE DIFFERENCE IN THE COMPOSITION OF BACTERIAL CELL WALL COMPOSITION Gram positive organisms: with thick peptidoglycan layer and teichoic acid cross linkages. Gram negative organisms : thin layer of peptidoglycan.

GENERAL RULES : 1. ALL COCCI ARE GRAM POSITIVE EXCEPT

Neisseria, Veillonella, and Branhamella

2.

ALL BACILLI ARE GRAM NEGATIVE EXCEPT

Bacillus, Clostridium, Mycobacterium, Corynebacterium,Nocardia, Erysipelothrix, Lactobacillus, and Listeria.

ALL COCCI ARE NON-MOTILE AND NONSPORE-FORMER ALL ENCAPSULATED ORGANISMS ARE NONMOTILE

3.

4.

5. Bacillus and Clostridium ARE SPORE FORMING ORGANISMS 6. THE HIGHER FORMS OF ORGANISM INCLUDING Actinomyces, Streptomyces, yeast and mold ARE GRAM POSITIVE 7. SPIRAL ORGANISMS ARE NOT STAINABLE EXCEPT FOR SOME WHICH ARE GRAM NEGATIVE.

GRAM POSITIVE BACILLI

GRAM NEGATIVE BACILLI

PRINCIPLE : USED FOR SUBSET OF BACTERIA WHOSE CELL WALL CONTAINS LONG CHAIN FATTY ACID. ACID FAST : ORGANISMS WITH MYCOLIC ACID IN THEIR CELL WALL. NON-ACID FAST : ORGANISMS WITHOUT MYCOLIC ACID IN THEIR CELL WALL.

REAGENTS : 1. Carbol fuchsin 2. Acid Alcohol 3. Methylene Blue or Malachite Green

METHODS : ZIEHL NEELSEN METHOD KINYOUN METHOD (phenol) PAPPENHEIMS METHOD ( alcohol/Rosolic acid) BAUMGARTENS METHOD (diluted alc.fuchsin) RHODAMINE AURAMINE METHOD

ACID FAST BACILLI

NON-ACID FAST ORGANISMS

A.

CAPSULAR STAINS
1. HISSS COPPER SULFATE METHOD
- CAPSULATED ORGANISMS APPEAR AS DARK PURPLE BODY WITH A FAINT BLUE CAPSULE AROUND IT

2. GINS METHOD - BACTERIA WILL BE STAINED BUT THE CAPSULE IS UNSTAINED WITH THEIR MARGIN DELINEATED BY THE INK. 3. WELCHS METHOD - CAPSULE STAINS PALE VIOLET 4. WADSWORTHS METHOD -BACTERIA ( BLUE ), Capsule ( PINKISH )

5. INDIA INK METHOD 6. MUIR METHOD 7. ANTHONYS METHOD

LOEFFLERS ALKALINE METHYLENE BLUE ( LAMB ) 2. NEISSER STAIN - BACILLI APPEAR ENTIRELY BROWN OR SHOW DARK BLUE ROUND BODY AT BOTH ENDS. 3. ALBERTS METHOD - GRANULES APPEAR BLUE-BLACK 4. LJUBINSKY METHOD
1.

1.

HEAT AND ACETIC ACID

2. -BACTERIA (BLUE), SPORES (RED) DORNER METHOD -BACTERIA ( ALMOST COLORLESS), SPORES (RED) WITH DARK GRAY BACKGROUND WIRTZ CONKLIN METHOD -SPORES ARE SEEN AS GREEN SPHERULES IN RED STAINED RODS OR DEBRIS. 10% NIGROSIN AND CARBOL FUCHSIN SCHAEFFER AND FULTON - SPORES APPEAR AS LIGHT GREEN

3.

4. 5.

1.

LEIFSON METHOD
2. 3. 4. 5. 6. - FLAGELLA ( RED ), BACTERIAL CELL ( BLUE ) SILVER STAIN FOR FLAGELLA FISHER CONN CASARES GILS METHOD LOFFLERS METHOD VAN ERMENGENS METHOD

* FLAGELLAR STAIN USED A SPECIAL MORDANT SUCH AS TANNIC ACID THAT SWELLS, COATS, FORMS A PRECIPITATE WITH THE FLAGELLA.

1.

BURRIS INDIA INK METHOD

- SPIROCHETES/BACTERIA (WHITE IN DARK FIELD) -USED FOR ORGANISMS WITH POOR STAINING PROPERTIES 2. NIGROSIN - USED FOR STUDYING SPIROCHETES 3. RELIEF STAIN ( DORNER ) - CELLS ARE UNSTAINED AGAINST THE DARK BACKGROUND.