Chapter XXXI: Anaphylaxis (1)

Dallera, in 1874, and a number of physiologists of that period, observed peculiar skin eruptions following the transfusion of blood, that is, the introduction of foreign proteins. In the years subsequent to the introduction of diphtheria antitoxin (1890) characteristic “serum rashes” were not infrequently reported, sometimes accompanied by more or less severe general symptoms and occasionally death--a train of phenomena to which the name “serum sickness” was later applied, since it was shown that it was the horse serum (foreign protein) that was the cause, and not the antitoxin itself. In 1898 Richet and Hericourt noticed that some of the dogs which they were attempting to immunize against toxic eel serum not only were not immunized but suffered even more severely after the second injection. They obtained similar results with an extract of mussels which contain a toxin. Richet gave the name “anaphylaxis” (“no protection”) to this phenomenon to distinguish it from immunity or prophylaxis (protection).

All the above-mentioned observations led to no special investigations as to their cause. In 1903, Arthus noticed abscess formation, necrosis and sloughing following several injections of horse serum in immediately adjacent parts of the skin in rabbits (“Arthus’ phenomenon”). Theobald Smith, in 1904, observed the death of guinea-pigs following properly spaced injections of horse serum. This subject was investigated by Otto and by Rosenau and Anderson in this country and about the same time von Pirquet and Schick were making a study of serum rashes mentioned above. The publications of these men led to a widespread study of the subject of injections of foreign proteins. It is now a well-established fact that the injection into an animal of a foreign protein--vegetable, animal or bacterial, simple or complex--followed by a second injection after a proper length of time leads to a series of symptoms indicating poisoning, which may be so severe as to cause the death of the animal. Richet’s term “anaphylaxis” has been applied to the condition of the animal following the first injection and indicates that it is in a condition of supersensitiveness for the protein in question. The animal is said to be “sensitized” for that protein.[25] The sensitization is specific since an animal injected with white of chicken’s egg reacts to a second injection of chicken’s egg only and not pigeon’s egg or blood serum or any other protein. The specific poisonous substance causing the symptoms has been called “anaphylotoxin” though what it is, is still a matter of investigation. It is evident that some sort of an antibody results from the first protein injected and that it is specific for its own antigen.

A period of ten days is usually the minimum time that must elapse between the first and second injections in guinea-pigs in order that a reaction may result, though a large primary dose requires much longer. If the second injection is made within less time no effect follows, and after three or more injections at intervals of about one week the animal fails to react at all, it has become “immune” to the protein. Furthermore, after an animal has been sensitized by one injection and has reacted to a second, then, if it does not die from the reaction, it fails to react to subsequent injections. In this latter case it is said to be “antianaphylactic.”

It must be remembered that proteins do not normally get into the circulation except by way of the alimentary tract. Here all proteins that are absorbed are first broken down to their constituent amino-acids, absorbed as such and these are built up into the proteins characteristic of the animal’s blood. Hence when protein as such gets into the blood it is a foreign substance to be disposed of. The blood contains proteolytic enzymes for certain proteins normally. It is also true that the body cells possess the property of digesting the proteins of the blood and building them up again into those which are characteristic of the cell. Hence it appears rational to assume that the foreign proteins act as stimuli to certain cells to produce more of the enzymes necessary to decompose them, so that they may be either built up into cell structure or eliminated as waste. If in this process of splitting up of protein a poison were produced, then the phenomena of “anaphylaxis” could be better understood. As a matter of fact Vaughan and his co-workers have shown that by artificially splitting up proteins from many different sources--animal, vegetable, pathogenic and saprophytic bacteria--a poison _is produced_ which appears to be the same in all cases and which causes the symptoms characteristic of anaphylaxis. On the basis of these facts it is seen that anaphylaxis is simply another variety of immunity. The _specific antibody_ in this case is an _enzyme_ which decomposes the protein instead of precipitating it. The enzyme must be specific for the protein since these differ in constitution. Vaughan even goes so far as to say that the poison is really the central ring common to all proteins and that they differ only in the lateral groups or side chains attached to this central nucleus. The action of the enzyme in this connection would be to split off the side chains, and since these are the specific parts of the protein, the enzyme must be specific for each protein. The pepsin of the gastric juice and the trypsin of the pancreas split the native proteins only to peptones. As is well known, these when injected in sufficient quantity give rise to poisonous symptoms, and will also give rise to anaphylaxis under properly spaced injections. They do not poison normally because they are split by the intestinal erepsin to amino-acids and absorbed as such. Whether Vaughan’s theory of protein structure is the true one or not remains to be demonstrated. It is not essential to the theory of anaphylaxis above outlined, _i.e._, a phenomenon due to the action of specific _antibodies_ which are enzymes. On physiological grounds this appears the most rational of the few explanations of anaphylaxis that have been offered and was taught by the author before he had read Vaughan’s theory along the same lines.

On the basis of the author’s theory the phenomena of protein immunity and antianaphylaxis may be explained in the following way which the author has not seen presented. The enzymes necessary to decompose the injected protein are present in certain cells and are formed in larger amount by those cells to meet the increased demand due to injection of an excess of protein. They are retained in the cell for a time at least. If a second dose of protein is given before the enzymes are excreted from the cells as waste, this is digested within the cells in the normal manner. If a third dose is given, the cells adapt themselves to this increased intracellular digestion and it thus becomes normal to them. Hence the _immunity_ is due to this increased intracellular digestion.

On the other hand, if the second injection is delayed long enough, then the _excess_ enzyme, but not all, is excreted from the cells and meets the second dose of protein in the blood stream and rapidly decomposes it there, so that more or less intoxication from the split products results. This uses up _excess_ enzyme, hence subsequent injections are not digested in the blood stream but within the cells as before. So that “antianaphylaxis” is dependent on the exhaustion of the excess enzyme in the blood, and the condition is _fundamentally_ the same as protein immunity, _i.e._, due to _intracellular_ digestion in each case.

As has been indicated “serum sickness” and sudden death following serum injections are probably due to a sensitization of the individual to the proteins of the horse in some unknown way. Probably hay fever urticarial rashes and idiosyncrasies following the ingestion of certain foods--strawberries, eggs, oysters, etc., are anaphylactic phenomena.

In medical practice the reaction is used as a means of diagnosis in certain diseases, such as the tuberculin test in tuberculosis, the mallein test in glanders. The individual or animal with tuberculosis becomes sensitized to certain proteins of the tubercle bacillus and when these proteins in the form of tuberculin are introduced into the body a reaction results, local or general, according to the method of introduction. The practical facts in connection with the tuberculin test are also in harmony with the author’s theory of anaphylaxis as above outlined. Milder cases of tuberculosis give more vigorous reactions because the intracellular enzymes are not used up rapidly enough since the products of the bacillus are secreted slowly in such cases. Hence excess of enzyme is free in the blood and the injection of the tuberculin meets it there and a vigorous reaction results. In old, far-advanced cases, no reaction occurs, because the enzymes are all used in decomposing the large amount of tuberculous protein constantly present in the blood. The fact that an animal which has once reacted fails to do so until several months afterward likewise depends on the fact that the _excess_ enzyme is used in the reaction and time must elapse for a further excess to accumulate.

The anaphylactic reaction has been made use of in the identification of various types of proteins and is of very great value since the reaction is so delicate, particularly when guinea-pigs are used as test animals. Wells has detected the 0.000,001 g. of protein by this test. It is evident that the test is applicable in medico-legal cases and in food examination and has been so used.

A TABULATION OF ANTIGENS AND ANTIBODIES AS AT PRESENT RECOGNIZED.

CLASS OF
ANTIGEN ANTIBODY ACTION OF ANTIBODY RECEPTOR

Toxin Antitoxin Combines with toxin and I.
hence prevents toxin
from uniting with a
cell and injuring it,
_i.e._, neutralizes toxin.

Enzyme Antienzyme Combines with enzyme I.
and thus prevents enzyme
from uniting
with anything else and
showing its action, _i.e._,
neutralizes enzyme.

Solution of Precipitin Unites with its antigen II.
protein and causes its precipitation
from solution.

Solution of ? Causes phenomenon of (?)
protein anaphylaxis(?)

Suspension of Agglutinin Unites with its antigen II.
cells causes its clumping together
and settling out
of suspension.

Suspension of Opsonin Unites with its antigen II.
cells and makes the cells (?)
more easily taken up
by phagocytes.

Suspension of Amboceptor Unites with its antigen III.
cells and also with complement
which latter then
dissolves the antigen.

Precipitin Antiprecipitin Neutralizes precipitin. I.

Agglutinin Antiagglutinin Neutralizes agglutinin. I.

Opsonin Antiopsonin Neutralizes opsonin. I.

Amboceptor Antiamboceptor Neutralizes amboceptor. I.
(two kinds)

Complement Anticomplement Neutralizes complement. I.

SUMMARY OF IMMUNITY AS APPLIED TO PROTECTION FROM DISEASE.

The discussion of “immunity problems” in the preceding chapters serves to show that protection from disease either as a condition natural to the animal or as an acquired state is dependent on certain properties of its body cells or fluids, or both. The actual factors so far as at present known may be summarized as follows:

1. _Antitoxins_ which neutralize true toxins; shown to exist for very few diseases.

2. _Cytolytic substances_ which destroy the invading organism: in reality two substances; amboceptor, which is specific, and complement, the real dissolving enzyme.

3. _Phagocytosis_ or the destruction of the invading organisms within the leukocytes.

4. _Opsonins_ which render the bacteria more readily taken up by the phagocytes.

5. _Enzymes_ other than complement possibly play a part in the destruction of some pathogenic organisms or their products. This remains to be more definitely established.

6. It is possible that in natural immunity there might be no receptors in the body cells to take up the organisms or their products, or the receptors might be present in certain cells but of a very low chemical affinity, so that combination does not occur. It is even highly probable that many substances formed by invading organisms which might injure specialized cells, such as those of glandular, nervous or muscle tissue, have a more rapid rate of reaction with, or a stronger affinity for, lower unspecialized cells, such as connective and lymphoid tissue, and unite with these so that their effects are not noticed.

The importance of these different, factors varies in different diseases and need not be considered in this connection.

The question “which of the body cells are engaged in the production of antibodies” is not uncommonly asked. On physiological grounds it would not seem reasonable that the highly specialized tissues above mentioned could take up this work, even though they are the ones which suffer the greatest injury in disease. Hence it is to be expected that the lower or unspecialized cells are the source, and it has been shown that the antibodies are produced by the phagocytes (though not entirely as Metchnikoff maintained), by lymphoid tissue generally, by the bone marrow and also by connective-tissue cells, though in varying degrees.

Since immunity depends on the activity of the body cells it is evident that one of the very best methods for avoiding infectious diseases is to keep these cells up to their highest state of efficiency, to keep in “good health.” Hence good health means not only _freedom from disease_ but also _protection against disease_.

LIST OF LABORATORY EXERCISES GIVEN IN CONNECTION WITH THE CLASS WORK INCLUDED IN THIS TEXT-BOOK.

Exercise 1. Cleaning glassware.

Exercise 2. Preparation of broth medium from meat juice.

Exercise 3. Preparation of gelatin medium from broth.

Exercise 4. Preparation of agar medium from broth.

Exercise 5. Potato tubes.

Exercise 6. Potato plates.

Exercise 7. Plain milk tubes.

Exercise 8. Litmus milk tubes.

Exercise 9. Sugar broth media.

Exercise 10. Blood-serum tubes.

Exercise 11. Inoculation of tubes. Action on complex proteins.

Exercise 12. Production of gas from carbohydrates.

Exercise 13. Production of indol.

Exercise 14. Reduction of nitrates.

Exercise 15. Chromogenesis: Illustrates nicely the variation with
environment.

Exercise 16. Enzyme production.

Exercise 17. Making of plate cultures; isolation in pure culture.

Exercise 18. Stain making and staining.

Exercise 19. Cell forms and cell groupings.

Exercise 20. Hanging drop slides.

Exercise 21. Staining of spores.

Exercise 22. Staining of acid-fast bacteria.

Exercise 23. Staining of capsules.

Exercise 24. Staining of metachromatic granules.

Exercise 25. Staining of flagella.

Exercise 26. Study of individual species.

Exercise 27. Determination of thermal death-point.

Exercise 28. Action of disinfectants on bacteria.

Exercise 29. Action of sunlight on bacteria.

DESCRIPTIVE CHART--SOCIETY OF AMERICAN BACTERIOLOGISTS.

_Prepared by Committee on Methods of Identification of Bacterial Species.--F. D. Chester, F. P. Gorham, Erwin F. Smith._

_Endorsed by the Society for general use at the Annual Meeting, December, 1907._

GLOSSARY OF TERMS.

AGAR HANGING BLOCK, a small block of nutrient agar cut from a pour plate, and placed on a cover-glass, the surface next the glass having been first touched with a loop from a young fluid culture or with a dilution from the same. It is examined upside down, the same as a hanging drop.

AMEBOID, assuming various shapes like an ameba.

AMORPHOUS, without visible differentiation in structure.

ARBORESCENT, a branched, tree-like growth.

BEADED, in stab or stroke, disjointed or semiconfluent colonies along the lines of inoculation.

BRIEF, a few days, a week.

BRITTLE, growth dry, friable under the platinum needle.

BULLATE, growth rising in convex prominences, like a blistered surface.

BUTYROUS, growth of a butter-like consistency.

CHAINS,
Short chains, composed of 2 to 8 elements.
Long chains, composed of more than 8 elements.

CILIATE, having fine, hair-like extensions, like cilia.

CLOUDY, said of fluid cultures which do not contain pseudozoogleæ.

COAGULATION,[22] the separation of casein from whey in milk. This may take place quickly or slowly, and as the result either of the formation of an acid or of a lab ferment.

CONTOURED, an irregular, smoothly undulating surface, like that of a relief map.

CONVEX surface, the segment of a circle, but flattened.

COPROPHYL, dung bacteria.

CORIACEOUS, growth tough, leathery, not yielding to the platinum needle.

CRATERIFORM, round, depressed, due to the liquefaction of the medium.

CRETACEOUS, growth opaque and white, chalky.

CURLED, composed of parallel chains in wavy strands, as in anthrax colonies.

DIASTASIC ACTION, same as DIASTATIC, conversion of starch into water-soluble substances by diastase.

ECHINULATE, in agar stroke a growth along line of inoculation, with toothed or pointed margins; in stab cultures growth beset with pointed outgrowths.

EFFUSE, growth thin, veily, unusually spreading.

ENTIRE, smooth, having a margin destitute of teeth or notches.

EROSE, border irregularly toothed.

FILAMENTOUS, growth composed of long, irregularly placed or interwoven filaments.

FILIFORM, in stroke or stab cultures a uniform growth along line of inoculation.

FIMBRIATE, border fringed with slender processes, larger than filaments.

FLOCCOSE, growth composed of short curved chains, variously oriented.

FLOCCULENT, said of fluids which contain pseudozoogleæ, _i.e._, small adherent masses of bacteria of various shapes and floating in the culture fluid.

FLUORESCENT, having one color by transmitted light and another by reflected light.

GRAM’S STAIN, a method of differential bleaching after gentian violet, methyl violet, etc. The + mark is to be given only when the bacteria are deep blue or remain blue after counter-staining with Bismarck brown.

GRUMOSE, clotted.

INFUNDIBULIFORM, form of a funnel or inverted cone.

IRIDESCENT, like mother-of-pearl. The effect of very thin films.

LACERATE, having the margin cut into irregular segments as if torn.

LOBATE, border deeply undulate, producing lobes (see _Undulate_).

LONG, many weeks, or months.

MAXIMUM TEMPERATURE, temperature above which growth does not take place.

MEDIUM, nutrient substance upon which bacteria are grown.

MEMBRANOUS, growth thin, coherent, like a membrane.

MINIMUM TEMPERATURE, temperature below which growth does not take place.

MYCELIOID, colonies having the radiately filamentous appearance of mold colonies.

NAPIFORM, liquefaction with the form of a turnip.

NITROGEN REQUIREMENTS, the necessary nitrogenous food. This is determined by adding to _nitrogen-free_ media the nitrogen compound to be tested.

OPALESCENT, resembling the color of an opal.

OPTIMUM TEMPERATURE, temperature at which growth is most rapid.

PELLICLE, in fluid bacterial growth forming either a continuous or an interrupted sheet over the fluid.

PEPTONIZED, said of curds dissolved by trypsin.

PERSISTENT, many weeks, or months.

PLUMOSE, a fleecy or feathery growth.

PSEUDOZOOGLEÆ, clumps of bacteria, not dissolving readily in water, arising from imperfect separation, or more or less fusion of the components, but not having the degree of compactness and gelatinization seen in zoogleæ.

PULVINATE, in the form of a cushion, decidedly convex.

PUNCTIFORM, very minute colonies, at the limit of natural vision.

RAPID, developing in twenty-four to forty-eight hours.

RAISED, growth thick, with abrupt or terraced edges.

RHIZOID, growth of an irregular branched or root-like character, as in _B. mycoides_.

RING, same as RIM, growth at the upper margin of a liquid culture, adhering more or less closely to the glass.

REPAND, wrinkled.

SACCATE, liquefaction the shape of an elongated sac, tubular, cylindrical.

SCUM, floating islands of bacteria, an interrupted pellicle or bacteria membrane.

SLOW, requiring five or six days or more for development.

SHORT, applied to time, a few days, a week.

SPORANGIA, cells containing endospores.

SPREADING, growth extending much beyond the line of inoculation, _i.e._, several millimetres or more.

STRATIFORM, liquefying to the walls of the tube at the top and then proceeding downward horizontally.

THERMAL DEATH-POINT, the degree of heat required to kill young fluid cultures of an organism exposed for ten minutes (in thin-walled test-tubes of a diameter not exceeding 20 mm.) in the thermal water-bath. The water must be kept agitated so that the temperature shall be uniform during the exposure.

TRANSIENT, a few days.

TURBID, cloudy with flocculent particles; cloudy plus flocculence.

UMBONATE, having a button-like, raised centre.

UNDULATE, border wavy, with shallow sinuses.

VERRUCOSE, growth wart-like, with wart-like prominences.

VERMIFORM-CONTOURED, growth like a mass of worms or intestinal coils.

VILLOUS, growth beset with hair-like extensions.

VISCID, growth follows the needle when touched and withdrawn, sediment on shaking rises as a coherent swirl.

ZOOGLEÆ, firm gelatinous masses of bacteria, one of the most typical examples of which is the _Streptococcus mesenterioides_ of sugar vats. (_Leuconostoc mesenterioides_), the bacterial chains being surrounded by an enormously thickened, firm covering inside of which there may be one or many groups of the bacteria.

NOTES.

(1) For decimal system of group numbers see Table I. This will be found useful as a quick method of showing close relationships inside the genus, but is not a sufficient characterization of any organism.

(2) The morphological characters shall be determined and described from growths obtained upon at least one solid medium (nutrient agar) and in at least one liquid medium (nutrient broth). Growths at 37° C. shall be in general not older than twenty-four to forty-eight hours, and growths at 20° C. not older than forty-eight to seventy-two hours. To secure uniformity in cultures, in all cases preliminary cultivation shall be practised as described in the revised Report of the Committee on Standard Methods of the Laboratory Section of the American Public Health Association, 1905.

(3) The observation of cultural and biochemical features shall cover a period of at least fifteen days and frequently longer, and shall be made according to the revised Standard Methods above referred to. All media shall be made according to the same Standard Methods.

(4) Gelatin stab cultures shall be held for six weeks to determine liquefaction.

(5) Ammonia and indol tests shall be made at end of tenth day, nitrite tests at end of fifth day.

(6) Titrate with N/20 NaOH, using phenolphthalein as an indicator; make titrations at same time from blank. The difference gives the amount of acid produced.

The titration should be done after boiling to drive off any CO₂ present in the culture.

(7) Generic nomenclature shall begin with the year 1872 (Cohn’s first important paper).

Species nomenclature shall begin with the year 1880 (Koch’s discovery of the pour plate method for the separation of organisms).

(8) Chromogenesis shall be recorded in standard color terms.

TABLE I.

A NUMERICAL SYSTEM OF RECORDING THE SALIENT CHARACTERS OF AN ORGANISM. (GROUP NUMBER.)

100 Endospores produced
200 Endospores not produced
10 Aërobic (strict)
20 Facultative anaërobic
30 Anaërobic (strict)
1 Gelatin liquefied
2 Gelatin not liquefied
0.1 Acid and gas from dextrose
0.2 Acid without gas from dextrose
0.3 No acid from dextrose
0.4 No growth with dextrose
0.01 Acid and gas from lactose
0.02 Acid without gas from lactose
0.03 No acid from lactose
0.04 No growth with lactose
0.001 Acid and gas from saccharose
0.002 Acid without gas from saccharose
0.003 No acid from saccharose
0.004 No growth with saccharose
0.0001 Nitrates reduced with evolution of gas
0.0002 Nitrates not reduced
0.0003 Nitrates reduced without gas formation
0.00001 Fluorescent
0.00002 Violet chromogens
0.00003 Blue chromogens
0.00004 Green chromogens
0.00005 Yellow chromogens
0.00006 Orange chromogens
0.00007 Red chromogens
0.00008 Brown chromogens
0.00009 Pink chromogens
0.00000 Non-chromogenics
0.000001 Diastasic action on potato starch, strong
0.000002 Diastasic action on potato starch, feeble
0.000003 Diastasic action on potato starch, absent
0.0000001 Acid and gas from glycerin
0.0000002 Acid without gas from glycerin
0.0000003 No acid from glycerin
0.0000004 No growth with glycerin

The genus according to the system of Migula is given its proper symbol which precedes the number thus:(7)

BACILLUS COLI (Esch.) Mig. becomes B. 222.111102
BACILLUS ALCALIGENES Petr. becomes B. 212.333102
PSEUDOMONAS CAMPESTRIS (Pam.) Sm. becomes Ps. 211.333151
BACTERIUM SUICIDA Mig. becomes Bact. 222.232103

Source............ Date of Isolation.............. Name........ Group No.(1)...............

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_.

{Orientation (grouping)............................
Agar {Chains (No. of elements)........................
Hanging-block {_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 {Orientation (grouping)........
Hanging-block {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.

+----------------------------------+---+---+---+---+---+---+---+---+
| | D | S | L | M | G | M | | |
| | e | a | a | a | l | a | | |
| | x | c | c | l | y | n | | |
| | t | c | t | t | c | n | | |
| 1. Fermentation-tubes containing | r | h | o | o | e | i | | |
| peptone-water or | o | a | s | s | r | t | | |
| sugar-tree bouillon and | s | r | e | e | i | | | |
| | e | o | | | n | | | |
| | | s | | | | | | |
| | | e | | | | | | |
+----------------------------------+---+---+---+---+---+---+---+---+
| 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. | | | | | | | | |
+----------------------------------+---+---+---+---+---+---+---+---+

2. Ammonia production, _feeble_, _moderate_, _strong_, _absent_,
_masked by acids_.

3. Nitrates in nitrate broth.

_Reduced_, _not reduced_.

Presence of nitrites...........ammonia..................

Presence of nitrates...........free nitrogen............

4. Indol production, _feeble_, _moderate_, _strong_.

5. Toleration of Acids, _great_, _medium_, _slight_.

_Acids tested_..............

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.

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.

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_.

Per cent. killed................

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:

+-----------+-------------+---+---+---+---+---+
| | | M | T | K | A | r |
| | | i | e | i | m | e |
| | | n | m | l | t | s |
| | | u | p | l | . | t |
| | | t | e | i | | r |
| | | e | r | n | r | a |
| | | s | a | g | e | i |
| Substance | Method used | | t | | q | n |
| | | | u | q | u | |
| | | | r | u | i | g |
| | | | e | a | r | r |
| | | | | n | e | o |
| | | | | t | d | w |
| | | | | i | | t |
| | | | | t | t | h |
| | | | | y | o | |
| | | | | | | |
+-----------+-------------+---+---+---+---+---+
| | | | | | | |
+-----------+-------------+---+---+---+---+---+
| | | | | | | |
+-----------+-------------+---+---+---+---+---+
| | | | | | | |
+-----------+-------------+---+---+---+---+---+
| | | | | | | |
+-----------+-------------+---+---+---+---+---+
| | | | | | | |
+-----------+-------------+---+---+---+---+---+
| | | | | | | |
+-----------+-------------+---+---+---+---+---+
| | | | | | | |
+-----------+-------------+---+---+---+---+---+
| | | | | | | |
+-----------+-------------+---+---+---+---+---+
| | | | | | | |
+-----------+-------------+---+---+---+---+---+

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.

+-------------------------------------+
| BRIEF CHARACTERIZATION. |
| |
| Mark + or 0, and when two terms |
| occur on a line erase the one which |
| does not apply unless both apply. |
| |
+--------------------------------+----+
| M | Diameter over 1µ |----|
| O | Chains, filaments |----|
| R | Endospores |----|
| P | Capsules |----|
| H | Zooglea, Pseudozooglea |----|
| O | Motile |----|
| L | Involution forms |----|
| O | Gram’s stain |----|
| G | |----|
| Y | |----|
|(2)| |----|
+---+----------------------------+----+
| C | B | Cloudy, turbid |----|
| U | r | Ring |----|
| L | o | Pellicle |----|
| T | t | Sediment |----|
| U | h | |----|
| R +-----+----------------------+----+
| A | A | Shining |----|
| L | g | Dull |----|
| | a | Wrinkled |----|
| F | r | Chromogenic |----|
| E +-----+----------------------+----+
| A | G | Round |----|
| T | e | Proteus-like |----|
| U | l. | Rhizoid |----|
| R | | Filamentous |----|
| E | P | Curled |----|
| S | l | |----|
|(3)| a | |----|
| | t | |----|
| | e | |----|
| +-----+----------------------+----+
| | G S | Surface growth |----|
| | e t | Needle growth |----|
| | l a | |----|
| | . b.| |----|
| +-----+----------------------+----+
| | P | Moderate, absent |----|
| | o | Abundant |----|
| | t | Discolored |----|
| | a | Starch destroyed |----|
| | t | |----|
| | o | |----|
| +-----+---------------------------+
| | Grows at 37° C. |----|
| | Grows in Cohn’s sol. |----|
| | Grows in Uschinsky’s sol. |----|
|---+-----+----------------------+----+
| B | L f | Gelatin(4) |----|
| I | i a | Blood-serum |----|
| O | q c | Casein |----|
| C | u t | |----|
| H | i i | |----|
| E | - o | |----|
| M | n | |----|
| I +-----+----------------------+----+
| C | M | Acid curd |----|
| A | i | Rennet curd |----|
| L | l | Casein peptonized |----|
| | k | |----|
| F +-----+----------------------+----+
| E | Indol(3) |----|
| A | Hydrogen sulphide |----|
| T | Ammonia(3) |----|
| U | Nitrates reduced(3) |----|
| R | Fluorescent |----|
| E | Luminous |----|
| S | |----|
+---+----------------------------+----+
| D | Animal pathogen, epizoon |----|
| I | Plant pathogen, epiphyte |----|
| S | Soil |----|
| T | Milk |----|
| R | Fresh water |----|
| I | Salt water |----|
| B | Sewage |----|
| U | Iron bacterium |----|
| T | Sulphur bacterium |----|
| I | |----|
| O | |----|
| N | |----|
+---+----------------------------+----+

FOOTNOTES.

[1] Sir H. A. Blake has called attention to the fact that the “mosquito theory” of malaria is mentioned in a Sanscrit manuscript of about the 6th century A.D.

[2] Myxomycetes excepted, and they are probably to be regarded as animals--Mycetozoa.

[3] Centralblatt f. Bakteriologie, etc. LXIII. 1 Abt. Orig. 1912, 4, idem LXVI. 1 Abt. Orig. 1912, 323.

[4] The pronunciation of this word according to English standards is kok-si; the continental pronunciation is kok-kee; the commonest American seems to be kok-ki. We prefer the latter since it is easier and more natural and should like to see it adopted. (Author.)

[5] With the possible exception of blue green algæ which have been found with bacteria in the above-mentioned hot springs. Seeds of many plants have been subjected to as low temperatures as those above-mentioned without apparent injury.

[6] It is popularly supposed that in canning fruit, vegetables, meats, etc., all the air must be removed, since the organisms which cause “spoiling” cannot grow in a vacuum. The existence of anaërobic and facultative anaërobic bacteria shows the fallacy of such beliefs.

[7] “By cellulose is understood a carbohydrate of the general formula C₆H₁₀O₅ not soluble in water, alcohol, ether, or dilute acids but soluble in an ammoniacal solution of copper oxide. It gives with iodine and sulphuric acid a blue color and with iodine zinc chloride a violet and yields dextrose on hydrolysis.”--H. Fischer.

[8] The sulphur bacteria are partially prototrophic for S; probably the iron bacteria also for Fe. Some few soil bacteria have been shown to be capable of utilizing free H, and it seems certain that the bacteria associated with the spontaneous heating of coal may oxidize free C. So far as known no elements other than these six are directly available to bacteria.

[9] Only a few kinds of bacteria so far as known are proto-autotrophic. The nitrous and nitric organisms of Winogradsky which are so essential in the soil, and which might have been the first of all organisms so far as their food is concerned, and some of the sulphur bacteria are examples.

[10] The term _pathogenic_ is also applied to certain non-parasitic saprophytic bacteria whose products cause disease conditions, as one of the organisms causing a type of food poisoning in man (_Clostridium botulinum_), which also probably causes “forage poisoning” in domestic animals.

[11] The term “fermentation” was originally used to denote the process which goes on in fruit juices or grain extracts when alcohol and gas are formed. Later it was extended to apply to the decomposition of almost any organic substance. In recent years the attempt has been made to give a chemical definition to the word by restricting its use to those changes in which by virtue of a “wandering” or rearrangement of the carbon atoms “new substances are formed which are not constitutents of the original molecule.” It may be doubted whether this restriction is justified or necessary. A definition is at present scarcely possible except when the qualifying adjective is included as “alcoholic fermentation,” “ammoniacal fermentation,” “lactic acid fermentation,” etc.

[12] See “Oil and Gas in Ohio,” Bownocker: Geological Survey of Ohio, Fourth Series, Bull. I, pp. 313-314.

[13] It is probable that this is the way “Jack o’lanterns” or “Will o’ the wisps” are ignited. Marsh gas is produced as above outlined from the vegetable and animal matter decomposing in swampy places under anaërobic conditions and likewise phosphine. These escape into the air and the “spontaneous combustion” of the phosphine ignites the marsh gas.

[14] Dr. H. H. Green, of Pretoria, South Africa, has isolated from “cattle dips” a bacterium that _reduces arsenates_ to _arsenites_.

[15] Dr. Green (l. c.) has also isolated an organism which causes some deterioration of cattle dips by _oxidizing arsenites to arsenates_.

[16] It will be noted that the names of enzymes (except some of those first discovered) terminate in _ase_ which is usually added to the _stem of the name of the substance acted on_, though sometimes to a word which indicates the substance formed by the action, as _lactacidase_, _alcoholase_.

[17] Tetanus toxin is about 120 times as poisonous as strychnin, both of which act on the same kind of nerve cells.

[18] In the author’s laboratory in the past ten years all sterilization except those few objects in blood and serum work which must be dry, has been done in autoclaves of the type shown in Fig. 81 which are supplied with steam from the University central heating plant. A very great saving of time is thus secured.

[19] The author has tested an “electric milk purifier” (Fig. 102) which was as efficient as a first-class pasteurizer and left the milk in excellent condition both chemically and as far as “cream line” was concerned. The cost of operation as compared with steam will depend on the price of electricity.

[20] The exact laboratory details for preparing various media are not given in this chapter. It is the object to explain the choice of different materials and the reasons for the various processes to which they are subjected.

[21] For a discussion of this method of standardization consult the following:

Clark & Lubs--J. Bact., 1917, II, 1-34, 109-136, 191-236.
Committee Report--Ibid., 1919. IV, 107-132.
Jones--J. Inf. Dis., 1919, 25, 262-268.
Fennel & Fisher--Ibid., 444-451.

Additional references will be found in these articles.

[22] Term also applied to the solidification of serum in media: _e.g._, the Hiss inulin medium for the differentiation of pneumococci (see diplococcus of pneumonia).

[23] The term “antigen” is also used to designate substances which may take the place of what are supposed to be the true antigens in certain diagnostic reactions (Chapter XXIX, Complement Fixation Test for Syphilis).

[24] If the antitoxin is later concentrated (see last paragraph in this chapter) a serum containing as little as 175 units per cc. may be commercially profitable.

[25] Tho term “allergie” was introduced by Von Pirquet to designate the state of the animal’s being sensitized and “allergic” as the adjective derived therefrom. It does not seem to the author that there is any advantage gained by the introduction of these terms.

INDEX

A

ABBÉ, 17
condenser, 200
microscope, improvements in, 30, 36

ABILGAARD, 26

Abrin, 262

Absorption of free nitrogen, 117
tests, 267

Accidental carriers, 241
structures, 43

Acetic acid, 99
bacteria, carbon oxidation, 114
fermentation, 32

_Acetobacter acidi oxalici_, 83
_xylinum_, 83

_Achorion schœnleinii_, 27, 34

Acid, acetic, 99
fermentation, 32
agglutination, 266
amino, relation to green plants, 119
butyric, 99
fermentation, 32, 99
carbolic, first used, 29
disinfectant action of, 159
fast bacteria, fat content, 84
staining of, 209
fermentation, 93
Bulgarian fermented milk, 98
ensilage, 98
industrial uses, 97
lactic acid, 96
sauerkraut, 98
hydrochloric, 246
production of, 110
soils, 81

Acquired immunity, 251, 252

_Actinomyces bovis_, 30, 36

Actinomycosis, cause of, 30, 36
path of entrance of, 244

Actions, reducing, 113

Activating enzymes, 125

Active immunity, definition of, 251, 252
production of, 252

Activities of bacteria, importance of, 31
overproduction, of cells, 258
physiological, definition of, 87
in identification, 216

Acute coryza, 244
disease, 233

Adulteration of food, anaphylactic test in, 293
complement-fixation test in, 279
immunity reactions in, 255
precipitin test in, 269

Aërobes, facultative, 76
strict, 76

Aërobic, 76, 215

Agar, composition of, 179
gelatinizing temperature, 179
medium, preparation of, 179
melting point of, 179
plating in, 188
sterilization of, 180

Agent, chemical, for disinfection, 156-163
choice of, for disinfection, 164
physical, for disinfection, 131

Agglutinating group, 266

Agglutination, acid, 266
diagnostic value of, 266
in identification of bacteria, 266
macroscopic, 265
microscopic, 265
phenomenon, 265

Agglutinin, 265
absorption test for, 267
action of, 266
anti-, 270
antigenic action of, 270
bacterial, 265
chief, 267
co-, 267
function of, 266
normal, 266
partial, 267
relation to precipitins, 269
specificity of, 267
theory of formation, 265
use of, 266

Agglutinogen, 266

Agglutinoid, 270

Aggressins, 288

Air, bacteria in, 71
filtration of, 153
“germ-free,” 153

Albumin in bacteria, 84

Alcohol as antiseptic, 160
as disinfectant, 160

Alcoholase, 125

Alcoholic fermentation, 31, 100

Alexin, 271, 273

Algæ, relation to bacteria, 37

Alimentary tract as path of entrance, 246

Alkalies as disinfectants, 158

Allergic, 290

Amboceptor, 273
anti-, 275
co-, 274
in cobra, 275
formation of, 273
hemolytic, 278
partial, 274
in rattle snake, 275
specificity of, 274
theory of formation, 273

Amboceptorogen, 274

Amebic dysentery, 29, 35

Ameboid cells, 247
colonies, 224

Amino-acids, relation to green plants, 119

Ammonia, structural formula, 103

Ammoniacal fermentation, 32

_Amœba coli_, 29, 35

Amphitrichic, 46

Amylase, 124

Anaërobes, 76
cultivation, methods of, 188
principles underlying, 188
facultative, 76
isolation of, 190
relation to elements, 86
strict, 76

Anaërobic, 76, 215
acid, butyric, 99
acid fermentation, 98
bacteria, first discovered, 32
fermentation of polysaccharides, 95

Analysis of ash, 82
chemical, of tubercle bacilli, 85

Anaphylactic, anti-, 290
phenomena, 292
reaction, uses of, 293

Anaphylatoxin, 290

Anaphylaxis, 289
anti-, 292
antibodies in, 291
theory of, 290, 291, 292

ANAXIMANDER, 18

ANDERSON, 289

ANDERSON and MCCLINTIC, phenol coefficient, 165

ANDRY, 25, 33

Anilin dyes, as antiseptic, 162
as disinfectants, 162
introduction of, 30
as stains, 204
Weigert, 36
fuchsin, 205
gentian violet, 205
water, 205

Animal carriers, 239
inoculation, uses of, 227

Animalcules, 19, 33

Animals, disinfection of, 170
experimental, 227
food relationships of, 39

_Ankylostoma duodenale_, discovery of, 27, 34
Egyptian chlorosis, cause of, 28, 35
hookworm disease, cause of, 28

Anthrax, 17, 28, 35
bacterium a facultative saprophyte, 238
isolation of, 29
due to a bacterium, 29
in human beings, 238
path of entrance, 243
intestine, 246
stomach, 246
persistence due to spores, 251
produced by exhaustion, 251
protective inoculation in, 30
spores, 29, 35
transmission by flies, 242
vaccine, 254

Anti-agglutinins, 270
aggressins, 288
amboceptors, 275
antisera in snake poisoning, 275
anaphylactic, 290
anaphylaxis due to intracellular digestion, 292
protein immunity compared to, 292
bacterial immunity, 254, 255
bodies, 259
place of production, 295
tabulation of, 294
body, action, 260
chemical composition, 260
formation of, 128, 260
complement, 274
complementophil amboceptor, 275
cytophil amboceptor, 275
diphtheritic serum, 263
enzyme, 122, 262
function of, 262

Antigen, 259
chemical composition of, 260
in complement-fixation, 277
syphilitic, 277, 279
in Wassermann test, 279

Antigens, fats and fatty acids as, 260
in preparation of vaccine, 285
tabulation of, 294

Antipollenin, 263

Antiprecipitins, 270

Antisepsis, 131
Lister, introduced, 35
primitive, 25

Antiseptic, 131
action of anilin dyes, 162
carbolic acid as, 159
cold as, 148

Antisera in snake poisoning, 275

Antisnake venoms, 275

Antitetanic serum, 263

Antitoxic immunity, 254, 255

Antitoxin, 261
collection of, 263
diphtheria, 30, 252
preparation of, 263
standard, 264
tetanus, 252

Antitoxins, 261-264
as factors in immunity, 295
preservative in, 263
specific, 261

Antivenin, 263

Apes, 227

Apparatus of Barber, 196

Appearance of growth on culture media, 217

APPERT, 20, 31, 34

Aqueous gentian violet, 205

Arborescent growth, 221

ARISTOTLE, 18

Aromatic compounds, production of, 104, 111

Arrak, 100

Arsenate, reduction of, 114

Arsenite, oxidation of, 115

ARTHUS, 289
phenomenon, 289

Articles, unwashable, disinfection of, 169
washable, disinfection of, 169

Artificial immunity, 251, 252

Ase, termination of name of enzyme, 124

Asepsis, 131

Aseptic, 131

Ash, analysis of, 82

Asiatic cholera, 27, 34, 73, 238, 239, 246, 248, 249

Attenuated, 253

Autoclave, air pressure sterilizer, 138
pressure sterilizer, 138

Autogenous vaccines, 284
in epidemic, 241

Autoinfection, 234

Autolysis, 149
self-digestion, 126

Autotrophic, 86

Available nitrogen, loss of, 113

Azotobacter, 118

B

BABES-ERNST corpuscles, 45

Bacilli, butter, 209
colon, 248
grass, 209
size and shape of, 52
tubercle, chemical analysis of, 85

Bacillus, 52, 60, 62
_anthracis_, 17, 36
spore staining, 209

Bacillus of blue milk, 31
Ducrey’s, 245
_subtilis_, 77, 83
spore staining, 209

Bacteria, absorption of N by, 117
acid fast, 84, 209
adaptability, range of, 90
advantage of motility to, 45
aids in isolation of, 197
anaërobic, 32
cause of disease in animals, 30
of souring of milk, 32
cell groupings of, 55
chains of, 38
chemical composition of, 39, 81
elements in, 82
classed as fungi, 37
as plants, 33, 35
definition of, 40
development of, 90
distribution of, 71
energy relationships, 39
environmental conditions for growth, 72
first classification of, 34
drawings of, 20
seen, 19, 33
food relationships of, 39
injurious, 72
isolation of, 194
measurement of, 40, 203
metabolism of, 86
methods of study of, 171
morphology of, 41
motile, 45
nitric, 114
nitrous, 114
nucleus of, 42
occurrence, 71
pathogenic, outside the body, 237
phosphorescent, 111, 112
position of, 37
rate of division, 43
of motion, 45
relation to algæ, 33, 37
to elements, 86
to gas and oil, 95
to phosphate rock, 115
to protozoa, 40
to soil fertility, 120
to sulphur deposits, 116
to yeasts and torulæ, 37
reproduction of, 37, 55
root tubercle, 86, 87
size of, 37, 40
soil, chief function of, 119
source of N, 102
speed of, 45
spiral, 53
staining of, 204-212
sulphur, 63
thermophil, 75, 77
universal distribution of, 90
in vinegar-making, 99

BACTERIACEÆ, 62, 66, 70

Bacterial agglutinin, 265
vaccines, 282
preparation of, 283, 284

Bacterin, 253

Bacteriocidin, 272

Bacteriological culture tubes, 184
examination, material for, 228
microscope, 200

Bacteriology, pathogenic, definition of, 231
reasons for study of, 217
as a science, 17, 32

Bacteriolysin, 272

Bacteriopurpurin, 62, 63, 112

Bacteriotropin, 281

_Bacterium abortus_, agglutinin of, 265
_coli_ in autoinfection, 234
gas formation by, 95
oxygen limits for, 77
pneumonia through intestinal route, 246
in preparation of sugar broths, 176
definition of, 62, 67, 70
_enteriditis_, cause of food poisoning, 104
_fluorescens_, oxygen limits, 77
_typhosum_, 73
agglutinin, 265
in phenol coefficient method, 166
pneumonia through intestinal route, 246

Ballon pipette, 193

Balsam, mounting in, 207

BARBER, 253
apparatus, 196

Barnyards, disinfection of, 167

Baskets, wire, 184

BASSI, 27
silkworm disease, 34

BASTIAN, 24

BAUMGÄRTNER, 256

Beaded growth, 221

Bed-bugs, 241

Beds, contact, 116
hot, 117

Beer, pasteurization of, 141, 144, 145

_Beggiatoa_, 63

BEGGIATOACEÆ, 63

BEHRING, 30

BELFANTI, 271

BERG, 27, 34

Berkefeld filter, 154

Bichloride of mercury as disinfectant, 158

BILHARZ, 28, 35

Bilharzia disease, 28, 35

Biochemical reactions, definition of, 87

Biological relationships, immunity reactions, 255, 270

Bipolar germination of spore, 48

Bismarck brown, 209, 212

Black-leg, 51, 73, 238, 243, 248, 251
vaccine, 254

Bleaching powder as disinfectant, 158

Blood, collection of, 228
cytolytic power of, 272
detection of, 269
serum, liquid, sterilization of, 182
Loeffler’s, 182
medium, preparation of, 182, 183
sterilization of, 182
vessels in dissemination of organisms, 247

Blue milk, bacterial cause of, 34
fermentation of, 31, 34

BOEHM, 27, 34

Boiling as disinfectant, 133

Boils, 237, 240, 243

BOLLINGER, 29, 30, 35, 36

BONNET, 20, 33

BORDET, 271

_Botrytis bassiana_, 27, 34

Bottles, staining of, 206

Bougies, 154

Bouillon, 173

BOYER, 260

Bread, salt rising, 95, 97

Bronchopneumonia, 233, 246

Broth, appearance of growth in, 218
extract of, 176
glycerine, 176
medium, 173
nitrate, 177
sterilization of, 174
sugar, 176

Brownian movement, 47, 203

Brushes, disinfection of, 169

Bubonic plague, 239

BUCHNER, 271

Budding of yeasts, 37

Bulgarian fermented milk, 98

Burning as disinfectant, 132

Burying as disinfectant, 154

BÜTSCHLI, 41, 43

Butter, 97
bacilli, staining of, 209
rancidity of, 101

Butyric acid fermentation, 32, 99

Buzzards, 241

C

CABBAGE disease due to protozoa, 36

Cadaverin, 104

CAIGNARD-LATOUR, 31, 34

Calcium hypochlorite as disinfectant, 158
oxide as disinfectant, 158

Candles, filter, 153, 154

Canned goods, food poisoning by, 104
spoilage of, 51

Canning, introduced, 21, 34
principles involved, 133

Capsule, 44, 45
of spore, 48
staining of, 210

Carbohydrates in bacterial cell, 84
fermentation of, 93-101

Carbol-fuchsin, 206

Carbolic acid as antiseptic, 159
as disinfectant, 159
first used, 29

Carbol-xylol, 209

Carbon cycle, 107
dioxide, 108
function of, in bacteria, 88, 101
oxidation of, 114
in proteins, liberation of, 105
source of, 88
uses of, 88, 101

CARBONI, 271

CARDANO, 18

Carrier problem, solution of, 240

Carriers, 239
accidental, 241
carrion eating animals as, 241
control of, 240
intermediate hosts as, 242
protective measures against, 242
universal, 240
of unknown organisms, 239

Cars, stock, disinfection of, 170

Catalase, 125

Catalytic agents, function of, 123

Catalyzer, 123

Cattle, 227

Causation of disease, 24, 128

Cell, constituents of, 84
contents of, 41, 83
forms of, 58, 59
staining for, 212
typical, 52
groupings, 55, 58, 59
staining for, 195
metabolism, 90
structures of, 41
wall, 41, 59
composition of, 83

Cells, chemical stimuli of, 257
overproduction activity of, 258
specific chemical stimuli of, 258

Cellular theory of immunity, 256, 280

Cellulose, definition of, 83
occurrence of, 83

Chain, 56

Channels of infection, 243
alimentary tract, 246
conjunctive, 244
external auditory meatus, 244
genitalia, 245
intestines, 246
lungs, 245
milk glands, 244
mouth cavity, 244
mucosæ, 244
nasal cavity, 244
pharynx, 245
skin, 243
stomach, 246
tonsils, 245

Chaos, 25

Characteristic groupings, 58

Characteristics of enzymes, 121
of toxins, 126

CHARRIN, 265

Chart, descriptive, 217

CHAUVEAU, 256

Cheese, eyes in, 96
failures, 110
Limburger, 101
odor of, 99
poisoning, 104
ripening of, 35

Chemical composition of bacteria, 39, 81, 85
elements in bacteria, 82
disinfectants, action of, 156-163
stimuli, 257-260
theory, fundamentals of, 256

Chemotherapy, 249, 255

CHEVREUIL, 21, 27, 31, 34

Chicken cholera, 30
pox, 239, 246

Chief agglutinin, 267
cell, 267

Chitin, 72

CHLAMYDOBACTERIACEÆ, 63

_Chlamydothrix_, 63

Chloride of lime as disinfectant, 158

Chlorine as disinfectant, 157

Chloroform as antiseptic, 162
as disinfectant, 162

Chlorophyl, 37, 112

Chlorosis, Egyptian, 27, 35

Cholera, Asiatic, carriers of, 239
organisms in, 27, 34
facultative saprophytes, 238
path of elimination of, 248
of entrance of, 246
relation to moisture, 73
specific location of, 249
hog, 242, 248, 252

Cholesterins as cell constituents, 84

Chromogenesis, 112

Chromoparic, 112

Chromophoric, 112

Chronic disease, 232

Chronological table, 33-36

Chymosin, 124

Circulation of carbon, 107
of nitrogen, 107
of phosphorus, 107
of sulphur, 108

Classification, advantage of, 59
early, 33, 35, 59
Migula’s, 62-63
S. A. B., 63-70

Cleaning of slides, 207

Clearing of sections, 209

Closed space disinfection, 161

_Clostridium_, 49
_botulinum_, 87, 104, 128, 238, 261
_pasteurianum_, 118
_tetani_, 128, 209, 233, 261, 263

Clothing, disinfection of, 170

Coagglutinins, 267

Coagulases, 124

Coagulating enzymes, 124

Coagulation temperature of proteins, 51

Coal, spontaneous heating of, 88

Coamboceptors, 274

Cobra, 275

COCCACEÆ, 62, 66, 68

Coccus, appearance of, on dividing, 57
cell form of, 52
groupings of, 52, 56, 57
division of, 52

Coenzymes, 122

COHN, 28, 33, 35, 59

Cold as antiseptic, 148
incubator, 215
storage, 148

Colds, due to universal carriers, 240
path of entrance of, 244
vaccines in, 241

Colonies, characteristics of plate, 223-226
definition of, 173

Color production, 112

Colorimetric method of standardization, 175

Combustion, spontaneous, 116

Commensal, 87

Commercial preparation of lactic acid, 99
products, why keep, 131
vaccines, 285

Communicable disease, 232

Complement, 273
deviation test, 277
effect of temperature on, 274
fixation test, 276-279
lecithin as, 274
relation to toxins and enzymes, 273
source of, 277

Complementoid, 274

Complementophil haptophore, 273

Complements, nature of, 274

Composition, chemical, 81-85
related to fungi, 39
relation to food, 81

Concentration of antitoxin, 264

Condenser, 200

Conditions for growth, general, 72
maximum, 72
minimum, 72
optimum, 72
spore formation, 51

Congenital immunity, 251, 252

Conjunctiva as path of entrance, 244

Constant temperature apparatus, 213

Contact beds, 116

Contagion, direct and indirect, 34