DETAILED FEATURES.

NOTE—Underscore required terms. Observe [notes] and [glossary of terms] on opposite side of card.

I. MORPHOLOGY[(2)]
- 1. Vegetative Cells,
  - Medium used............................. temp....................age.................days
  - Form, round, short rods, long rods, short chains, long chains, filaments, commas, short spirals, long spirals, clostridium, cuneate, clavate, curved.
  - Limits of Size.............................
  - Size of Majority.............................
  - Ends, rounded, truncate, concave.
  - Agar Hanging-block
  - Orientation (grouping)...............
  - Chains (No. of elements)...............
  - Short chains, long chains
  - Orientation of chains, parallel, irregular.
- 2. Sporangia,
  - medium used............................. temp....................age....................days
  - Form, elliptical, short rods, spindled, clavate, drumsticks.
  - Limits of Size.............................
  - Size of Majority.............................
  - Agar Hanging-block
  - Orientation (grouping)...............
  - Chains (No. of elements)...............
  - Orientation of chains, parallel, irregular.
  - Location of Endospores, central, polar.
- 3. Endospores.
  - Form, round, elliptical, elongated.
  - Limits of Size.............................
  - Size of Majority.............................
  - Wall, thick, thin.
  - Sporangium wall, adherent, not adherent.
  - Germination, equatorial, oblique, polar, bipolar, by stretching.
- 4. Flagella, No...............Attachment polar, bipolar, peritrichiate. How Stained...............
- 5. Capsules, present on...............
- 6. Zooglea, Pseudozooglea.
- 7. Involution Forms, on............in............days at............° C.
- 8. Staining Reactions.
  - 1:10 watery fuchsin, gentian violet, carbol-fuchsin, Loeffler’s alkaline methylene blue.
  - Special Stains.
    - Gram........................
    - Glycogen........................
    - Fat........................
    - Acid-fast........................
    - Neisser........................
II. CULTURAL FEATURES[(3)]
- 1. Agar Stroke.
  - Growth, invisible, scanty, moderate, abundant.
  - Form of growth, filiform, echinulate, beaded, spreading, plumose, arborescent, rhizoid.
  - Elevation of growth, flat, effuse, raised, convex.
  - Lustre, glistening, dull, cretaceous.
  - Topography, smooth, contoured, rugose, verrucose.
  - Optical characters, opaque, translucent, opalescent, iridescent.
  - Chromogenesis[(3)]........................
  - Odor, absent, decided, resembling........................
  - Consistency, slimy, butyrous, viscid, membranous, coriaceous, brittle.
  - Medium grayed, browned, reddened, blued, greened.
- 2. Potato.
  - Growth scanty, moderate, abundant, transient, persistent.
  - Form of growth, filiform, echinulate, beaded, spreading, plumose, arborescent, rhizoid.
  - Elevation of growth, flat, effuse, raised, convex.
  - Lustre, glistening, dull, cretaceous.
  - Topography, smooth, contoured, rugose, verrucose.
  - Chromogenesis[(3)]........................Pigment in water insoluble, soluble: other solvents................................................
  - Odor, absent, decided, resembling........................
  - Consistency, slimy, butyrous, viscid, membranous, coriaceous, brittle.
  - Medium, grayed, browned, reddened, blued, greened.
- 3. Loeffler’s Blood-serum.
  - Stroke invisible, scanty, moderate, abundant.
  - Form of growth, filiform, echinulate, beaded, spreading, plumose, arborescent, rhizoid.
  - Elevation of growth, flat, effuse, raised, convex.
  - Lustre, glistening, dull, cretaceous.
  - Topography, smooth, contoured, rugose, verrucose.
  - Chromogenesis[(3)]..........................
  - Medium grayed, browned, reddened, blued, greened.
  - Liquefaction begins in...............d, complete in...............d,
- 4. Agar Stab.
  - Growth uniform, best at top, best at bottom: surface growth scanty, abundant: restricted, wide-spread.
  - Line of puncture, filiform, beaded, papillate, villous, plumose, arborescent: liquefaction.
- 5. Gelatin Stab.
  - Growth uniform, best at top, best at bottom.
  - Line of puncture, filiform, beaded, papillate, villous, plumose, arborescent.
  - Liquefaction crateriform, napiform, infundibuliform, saccate, stratiform: begins in...............d. complete in...............d
  - Medium fluorescent, browned...............
- 6. Nutrient Broth.
  - Surface growth, ring, pellicle, flocculent, membranous, none.
  - Clouding slight, moderate, strong: transient, persistent: none: fluid turbid.
  - Odor, absent, decided, resembling..................
  - Sediment, compact, flocculent, granular, flaky, viscid on agitation, abundant, scant.
- 7. Milk.
  - Clearing without coagulation.
  - Coagulation prompt, delayed, absent.
  - Extrusion of whey begins in................days.
  - Coagulum slowly peptonized, rapidly peptonized.
  - Peptonization begins on...............d, complete on...............d.
  - Reaction, 1d......, 2d......, 4d......, 10d......, 20d......
  - Consistency, slimy, viscid, unchanged.
  - Medium browned, reddened, blued, greened.
  - Lab ferment, present, absent.
- 8. Litmus Milk.
  - Acid, alkaline, acid then alkaline, no change.
  - Prompt reduction, no reduction, partial slow reduction.
- 9. Gelatin Colonies.
  - Growth slow, rapid.
  - Form, punctiform, round, irregular, ameboid, mycelioid, filamentous, rhizoid.
  - Elevation, flat, effuse, raised, convex, pulvinate, crateriform (liquefying).
  - Edge, entire, undulate, lobate, erose, lacerate, fimbriate, filamentous, floccose, curled.
  - Liquefaction, cup, saucer, spreading.
- 10. Agar Colonies.
  - Growth slow, rapid (temperature................)
  - Form, punctiform, round, irregular, ameboid, mycelioid, filamentous, rhizoid.
  - Surface smooth, rough, concentrically ringed, radiate, striate.
  - Elevation, flat, effuse, raised, convex, pulvinate, umbonate.
  - Edge, entire, undulate, lobate, erose, lacerate, fimbriate, floccose, curled.
  - Internal structure, amorphous, finely, coarsely granular, grumose, filamentous, floccose, curled.
- 11. Starch Jelly.
  - Growth, scanty, copious.
  - Diastatic action, absent, feeble, profound.
  - Medium stained.......................
- 12. Silicate Jelly (Fermi’s Solution).
  - Growth copious, scanty, absent.
  - Medium stained.......................
- 13. Cohn’s Solution.
  - Growth copious, scanty, absent.
  - Medium fluorescent, non-fluorescent.
- 14. Uschinsky’s Solution.
  - Growth copious, scanty, absent.
  - Fluid viscid, not viscid.
- 15. Sodium Chloride in Bouillon.
  - Per cent. inhibiting growth........................
- 16. Growth in Bouillon over Chloroform, unrestrained, feeble, absent.
- 17. Nitrogen. Obtained from peptone, asparagin, glycocoll, urea, ammonia salts, nitrogen.
- 18. Best media for long-continued growth
- ..............................................................................................
- 19. Quick tests for differential purposes
- ..............................................................................................
  ..........................................................................................
III. PHYSICAL AND BIOCHEMICAL FEATURES.
- 1. Fermentation-tubes containing peptone-water or sugar-tree bouillon and
- Dextrose
- Saccharose
  - Reduced, not reduced.
  - Presence of nitrites..................ammonia..................
  - Presence of nitrates..................free nitrogen..................
- Lactose
- Maltose
  - Acids tested..................
- Glycerin
- Mannit
- Gas production, in per cent.
- (H/CO₂)
  - Thermal death-point (10
  - minutes’ exposure in nutrient broth when this is adapted to growth of organism)..........C.
  - Optimum temperature for growth..........° C.; or best growth at 16° C., 20° C., 25° C., 30° C., 37° C., 40° C., 50° C., 60° C.
  - Maximum temperature for growth..........° C.
  - Minimum temperature for growth..........° C.
- Growth in closed arm
- Amount of acid produced 1d.
- Amount of acid produced 2d.
  - Per cent. killed................
- Amount of acid produced 3d.
- 2. Ammonia production, feeble, moderate, strong, absent, masked by acids.
- 3. Nitrates in nitrate broth.
- 4. Indol production, feeble, moderate, strong.
- 5. Toleration of Acids, great, medium, slight.
- 6. Toleration of NaOH, great, medium, slight.
- 7. Optimum reaction for growth in bouillon, stated in terms of Fuller’s scale..........................
- 8. Vitality on culture media, brief, moderate, long.
- 9. Temperature relations.
- 10. Killed readily by drying: resistant to drying.
- 11. Per cent. killed by freezing (salt and crushed ice or liquid air)................
- 12. Sunlight: Exposure on ice in thinly sown agar plates; one-half plate covered (time 15 minutes), sensitive, not sensitive.
- 13. Acids produced................
- 14. Alkalies produced................
- 15. Alcohols................
- 16. Ferments, pepsin, trypsin, diastase, invertase, pectase, cytase, tyrosinase, oxidase, peroxidase, lipase, catalase, glucase, galactase, lab, etc.........................
- 17. Crystals formed:.........
- 18. Effect of germicides:
- Substance
- Method used
- Minutes
- Temperature
- Killing quantity
- Amt. required to restrain growth
IV. PATHOGENICITY.
- 1. Pathogenic to Animals.
  - Insects, crustaceans, fishes, reptiles, birds, mice, rats, guinea-pigs, rabbits, dogs, cats, sheep, goats, cattle, horses, monkeys, man..........................
- 2. Pathogenic to Plants:
- .............................................................
  .........................................................
  .........................................................
- 3. Toxins, soluble, endotoxins.
- 4. Non-toxin forming.
- 5. Immunity bactericidal.
- 6. Immunity non-bactericidal.
- 7. Loss of virulence on culture-media: prompt, gradual, not observed in.....................months.

Agar Hanging-block	Orientation (grouping)...............
	Chains (No. of elements)...............
	Short chains, long chains
	Orientation of chains, parallel, irregular.

Agar Hanging-block	Orientation (grouping)...............
	Chains (No. of elements)...............
	Orientation of chains, parallel, irregular.

1. Fermentation-tubes containing peptone-water or sugar-tree bouillon and	Dextrose	Saccharose	Lactose	Maltose	Glycerin	Mannit
Gas production, in per cent.
(H/CO₂)
Growth in closed arm
Amount of acid produced 1d.
Amount of acid produced 2d.
Amount of acid produced 3d.

Substance	Method used	Minutes	Temperature	Killing quantity	Amt. required to restrain growth

BRIEF CHARACTERIZATION. Mark + or 0, and when two terms occur on a line erase the one which does not apply unless both apply.
MORPHOLOGY[(2)]	Diameter over 1µ
	Chains, filaments
	Endospores
	Capsules
	Zooglea, Pseudozooglea
	Motile
	Involution forms
	Gram’s stain
CULTURAL FEATURES[(3)]	Broth	Cloudy, turbid
		Ring
		Pellicle
		Sediment
	Agar	Shining
		Dull
		Wrinkled
		Chromogenic
	Gel. Plate	Round
		Proteus-like
		Rhizoid
		Filamentous
		Curled
	Gel. Stab.	Surface growth
	Gel. Stab.	Needle growth
	Potato	Moderate, absent
		Abundant
		Discolored
		Starch destroyed
	Grows at 37° C.
	Grows in Cohn’s sol.
	Grows in Uschinsky’s sol.
BIOCHEMICAL FEATURES	Liquifaction	Gelatin[(4)]
		Blood-serum
		Casein
	Milk	Acid curd
		Rennet curd
		Casein peptonized
	Indol[(3)]
	Hydrogen sulphide
	Ammonia[(3)]
	Nitrates reduced[(3)]
	Fluorescent
	Luminous
DISTRIBUTION	Animal pathogen, epizoon
	Plant pathogen, epiphyte
	Soil
	Milk
	Fresh water
	Salt water
	Sewage
	Iron bacterium
	Sulphur bacterium