Chapter II: Part 2

In her experiments Doctor Day used the leaves, roots, and stems of the plants gathered in September. She believed that the greatest amount of poison is present in autumn and winter. She later undertook the isolation of the active principle, and proceeded as follows:[105]

The roots, stems, and leaves were boiled ten hours, strained, and
the decoction concentrated to a sirup, poured, while hot, into a
hot flask, corked and set away. At the end of ten days the sirup
had separated into two layers--the upper a blackish liquid, the
lower a brownish sediment. The liquid was poured into a flask and
covered with six times its volume of very dilute alcohol, 30 per
cent (the sediment also was washed with dilute alcohol, to insure a
complete removal of the liquid), corked, and let stand three days;
agitated occasionally, then filtered, and the filtrate slowly
evaporated in the air, when crystals were formed. It was found
important not to hurry the evaporation, for when this took place
too rapidly the crystals did not form.

These crystals are microscopic in size, blue-white in color, and of
a variety of forms. The most characteristic are slender and
pointed, arranged in rosettes or grouped in various ways. They are
soluble in distilled water and very dilute alcohol, very sparingly
soluble in strong alcohol, not soluble in chloroform or ether.

The evaporated mass containing the crystals, when dissolved in
distilled water, is slightly acid in reaction. A small amount of
this fed to a kitten produced the train of characteristic toxic
symptoms--sleepiness, loss of appetite, retching, and
diarrhea--that is produced by quite large amounts of the decoction.

The crystals Sayre[106] claims to have already seen. He says that they gave no precipitate with Mayer's reagent, platinum chlorid, or with ammonia, but that barium chlorid and ammonium oxalate gave a precipitate, and he believes that they were in reality an inorganic combination of calcium, so that while Doctor Day may have obtained an extract which produced characteristic symptoms she certainly has not isolated any pure active principle. Later she admitted that it was not possible "to make positive statements as to the chemical character of the active principle."[107]

In 1884 there was a fatal outbreak of a disorder in horses in portions of the Missouri Valley in Iowa, Nebraska, and Dakota. This was almost uniformly fatal in a few weeks or months. The animals lost strength and became emaciated, although they were kept in pasture where there was abundant grass. There was marked stupor, the animals falling asleep while eating, and they "would remain standing for a whole week, sleeping much of the time, with the head resting upon some object." The post-mortem examination showed that "in every instance there was marked hemorrhagic effusion into the fourth ventricle, the liver and spleen were abnormally dense, the walls of the intestines were almost destitute of blood, and the stomach enormously distended with undigested food." The post-mortem find and clinical symptoms suggested to Stalker[108] that this disorder was due to some plant analogous to _Astragalus mollissimus_. He found abundant in these regions _Crotalaria sagittalis_, or rattle-box, one of the so-called loco weeds, and by the administration per os to a young horse of an infusion of 15 pounds of the plant, given in two days, produced the clinical symptoms and the post-mortem condition of the brain which he previously observed on the range.

Power and Cambier[109] undertook the chemical study and the isolation of the active principle of this plant, together with that of _Astragalus mollissimus_. They found that the _Astragalus mollissimus_ if distilled with water yielded a distillate which possessed a peculiar odor, which they thought due to a trace of volatile oil. On distilling with alkali they obtained ammonia and a trace of trimethylamine. In the case of Crotalaria only ammonia was found.[110] They argued that because trimethylamine was not obtained in this case choline was not present. On distilling the _Astragalus mollissimus_ with acidulated water (H_{2}SO_{4}) the distillate was found to contain acetic acid--settling the nature of the "peculiar organic acid" described by Kennedy. From this plant they obtained a resin or mixture of resinous bodies by extracting the plant with alcohol, and after concentration precipitating with acid water. These resins in doses of from 2 to 5 grains failed to produce any symptoms in kittens.

An albuminoid which was obtained by precipitating a concentrated aqueous extract of _Astragalus mollissimus_ by means of alcohol likewise was found to be inactive to a kitten in doses corresponding to 50 grams of the crude plant. A globulin which was isolated by precipitation from a 10 per cent sodium chlorid solution proved also to be inactive in doses of 0.2 gram. They then extracted 3 kilograms of these plants with 1/2 per cent sulphuric acid, and after evaporation to a thick gum the mass was extracted with strong alcohol, the alcoholic solution was evaporated, and the alcoholic residue taken up in water and precipitated by neutral and basic lead acetates, and after removing the lead with sulphureted hydrogen the filtrate gave precipitates with various alkaloidal reagents. The sirupy residue which they obtained from _Astragalus mollissimus_ by decomposing the precipitate with Mayer's solution administered to kittens in doses of 0.1 gram produced merely frothing at the mouth with profuse flow of saliva, but the animals soon recovered. The presence of a large amount of calcium was shown but not estimated quantitatively.

Power and Cambier summed up their conclusions by stating that both the Astragalus and the Crotalaria contain very small amounts of toxic alkaloids, to which they believe the symptoms of poisoning produced were due. Their work from a chemical standpoint is excellent, but from a pharmacological point of view seems to be deficient; in fact, Power does not claim to be a pharmacologist. What would seem to be the proper course would have been to test for themselves the action of the plant on various animals and, after deciding which reacted most characteristically, test, after various precipitations, both the precipitates and filtrates on various animals to see whether the original symptoms and pathological lesions could be produced. They failed, however, to test their mother substance. It is well recognized that plants grown under varying conditions and on different soils vary in the amount of the physiologically active principle they contain.

In the case of Crotalaria, Power and Cambier had before them the experiment of Stalker, in which he reproduced the disorder by feeding the plant extract to horses, yet they claimed that the body which they administered was the active principle, merely because it produced some frothing at the mouth and salivation in a kitten. The percentage of active principle they found would be too small to account for the symptoms, except in the case of a very active compound.

Certain of these precipitates were also later examined physiologically by O'Brine.[111] He also found the resin precipitated from an alcoholic extract of the plant and also the alcoholic extract from 2.2 pounds of the dried _Astragalus mollissimus_ to be physiologically inactive.

Oatman,[112] using Power and Cambier's method with alfalfa (_Medicago sativa_), obtained a noncrystalline mass which when given in 0.1 gram dose caused frothing at the mouth in a kitten, but no serious symptoms. This 0.1 gram represented about 5 pounds of powdered leaves and tops of the plants.

Since the appearance of Power and Cambier's work Sayre has published various papers on the loco weeds in the Transactions of the Kansas Academy of Sciences for 1903-4, vol. 19, p. 194, 1905; 1901-2, vol. 18, p. 141; Seventh Biennial Report of the State Board of Agriculture of Kansas, vol. 12, p. 97, 1891; Journal of the Kansas Medical Society, vol. 4, pp. 222 and 241, 1904, etc. He has contributed nothing especially new, but says that "the old theory that an alkaloidal poison is secreted in the plant causing the loco trouble has not been found tenable," but wishes to be understood that he does not discredit the ground for the opinion that in some mysterious way certain disorders occur in cattle in connection with what is commonly called loco-weed. He suggests that this connection might be somewhat similar to the relationship between the disorder caused by over-feeding half-starved animals on clover or alfalfa[113] and has had the plant analyzed as to its nutritive value, giving the table in the Transactions of the Kansas Academy of Sciences, vol. 19, p. 194. He makes the suggestion that any injurious action the plants may have might be due to the fine, hair-like projections on the plant which mechanically set up irritation. This supposition can be thrown out at once by the experiment of Day and others, who induced symptoms in animals by extracts of the plant, and by the fact that other coarse plants do not act similarly. This fine, hair-like material was found to constitute about 33 per cent of the plant on grinding. But Sayre himself does not seem to be positive as to any conclusion. He, like O'Brine and others, has obtained alkaloidal reactions from the plant, but states he has obtained similar ones from alfalfa.[114] At one time he said:

I do not consider loco directly or indirectly the cause of the
condition, but am of the opinion that what is called "locoed" is,
first, congestion of the brain and spinal marrow (causing blindness
and first symptoms), and, second, softening to a greater or less
extent.[115] These terms describing the alleged symptoms of
"locoism" might occur in well recognized diseases resulting from
brain lesions, which latter occur in so-called forage poisoning and
poisoning from foul drinking water, etc.

We are not prepared to affirm or deny that the loco-weed produces a
train of symptoms characteristic of the plant.[116]

Again Sayre states:

It seems not unreasonable to suppose that the peculiar condition of
the animals of the plains, when they gorge themselves with this
highly nitrogenous weed, has something to do with the disease. A
condition of malnutrition may set in and give rise to the rapid
growth of a toxic-producing micro-organism or an irritating
principle. This principle may be capable of cultivation and of
producing disease artificially. Be this as it may, we feel
warranted in saying that the so-called poison is a development
within the animal, not a product preexisting in the weed itself.

Sayre also suggests the possibility of the plants producing hydro-cyanic acid, which, it is well known, occurs in sorghum.[117] In the Journal of the Kansas Medical Society (vol. 4, p. 243), he claims to have isolated a crystalline body, but this he has not tested physiologically. Sayre especially deserves credit for keeping the loco investigation alive, and no doubt his change in position is due to his lack of facilities for pharmacological testing.

Carl Ruedi[11] fed rabbits daily by a stomach tube with 10 c.c. of an extract (unstated strength) of _Astragalus mollissimus_ and recorded the following results:

After only five injections one of the rabbits died, and the
post-mortem showed to a nicety the congestion of the whole tract of
the vena portæ and the anæmia of the brain. I put six rabbits under
the influence of loco, and the effect was marked, but not rapid, if
not given in very concentrated solutions. The solutions were
prepared differently, and each of the rabbits had its own
preparation, but the effect was nearly the same. In the beginning
loco acts as a stimulant; the animals get lively, hilarious,
running about; cleaning themselves, etc. This lasts about eight
hours, then they become very quiet, sit in a corner of a box, and
one can do with them pretty nearly what one likes; they do not move
from the place, or just run into another corner, to fall back into
the same complacent reverie. One can leave the door open and hammer
away at the box, but they do not show any inclination to run away.
During the excitement, however, they become fierce, and I had once
the opportunity to watch one of the drollest things possible: One
of the rabbits, two hours after dosing it, got loose and ran under
a porch. A heavy tomcat came near this hole, and commenced sniffing
about; this offended the rabbit highly, and it jumped on the neck
of the cat, bit it through the skin, and the cat ran screaming
away. When the animals are first under the influence of moderate
doses of loco, they suffer greatly from hyperæsthesia of the
cutaneous nerves; when one touches them with a stick while lying in
a corner, without hurting them, one sees the platysma working away
very forcibly, and sometimes they utter sounds of pain. According
to my experiments the loco-weed works slowly but surely; as soon as
the anæmia of the brain sets in, the animals act in every respect
mad like; one hour they are excitable, and then again dull and
languid as can be. The rabbits eat, when well, very quickly, and
whenever they have opportunity; not so the locoed rabbit; he eats
slowly for a minute or two, then he goes into a corner and
meditates, comes forward to nibble at a carrot or a piece of
cabbage, but he never eats greedily, and does not steal it from the
mouth of his neighbor, or only very exceptionally. I observed these
rabbits for ten days; they did not die, because I gave them weaker
solutions; but they all became very ill, and as I had to leave the
park I killed them with the needle inserted into the medulla
oblongata, and made the post-mortem. In all of these cases I found
great congestion in the abdomen, and marked anæmia of the brain.
The congestion of the vena portæ commences certainly very early,
but still the first symptoms are the nervous symptoms, first as
excitants, then depressing or sedative, with a marked hyperæsthesia
of the cutaneous nerves.

Ruedi made an attempt to isolate the active principle and separated a base, which he calls "locoin," from an ether shaking. This base, however, he found to be physiologically inactive, but believes the activity to be due to a body which he calls "loco-acid," which is present in the mother liquid after the shaking with ether. He, however, has not obtained this in any degree of purity and gives no chemical data to substantiate this statement save that the fluid was acid.

Experiments made at the University of Pennsylvania with certain loco plants on cats, dogs, and rabbits proved negative.[119]

Other experiments on rabbits have been made by Doctor Lewis. These rabbits were fed on the leaves, stem, and whole plant, and also extracts of one of the loco plants (presumably _Astragalus mollissimus_) for one or two months, without producing any noticeable effect.[120]

This uncertainty in the results of the investigation as to the cause of the loco disease turned the attention of observers into other lines. President Ingersoll,[121] of the State Agricultural College of Colorado, in his autopsies on sheep was struck by the presence of tapeworms (_Taenia expansa_) in the gall duct and small intestines. He apparently tried to prove a relationship between the tapeworms and the locoed condition by feeding the extract of a loco plant to sheep, and thus showing its harmlessness. He prepared a decoction from 20 pounds of loco plant (the species was not stated) and boiled this down from 12 gallons to 1 quart. This concentrated extract was fed in three days to a bottle-fed lamb; this lamb showed no symptoms, although kept under observation for two weeks. This theory of the causation of loco by worms was also considered by Curtice,[122] and later brought forward by Steele[123] and Marshall.[124] This idea is very suggestive when considered in relation to the etiology of bothriocephalous anæmia.[125]

Others, again, have claimed that the disease is due to a parasite found upon the loco plants, but all specimens examined by entomologists proved to be harmless.[126]

Lloyd, from his study of the subject, says:

From first to last I have failed in obtaining a characteristic
proximate principle, either from the fresh or dried plant. The
disease called loco was as murky as the milk sickness so prevalent
in the new settlements of Indiana and Kentucky in early days, and,
like the numberless herbs that have been presumed to produce that
obscure peculiar disease, milk sickness, loco was unresponsive to
my chemistry.[127]

It may be safely said that if a specimen of the plant were to be
examined in the ordinary manner by a chemist who had no idea of its
importance he would report that it did not contain a characteristic
proximate constituent.[128]

Can it be that an admixture of loco and some undetermined plant or
earth infected with bacteria taken with the roots, each innocuous
under other conditions, can by digestion together in the stomach
and intestines result in the production of a poison?[129]

To sum up, it seems to the writer that the poison of loco is a
product, and not an educt.[130]

But Lloyd adds, in speaking of the reports of various experts and ranchmen:

Their description concerning its toxic action on animals agreed,
and it was folly to argue that so many observers from so many
sections of the country could be misled. There must be an
undetermined something behind the loco-weed.[131]

In 1893 O'Brine, from Colorado, and Mayo, from Kansas, reported on their work with the loco plants. O'Brine failed to isolate any alkaloidal or other poisonous body, and his feeding experiments on himself and on rabbits having failed, he sums up in despair: "The more I examine the loco question, the more I am persuaded that we must look for some other cause besides the loco-weed."[132] At the end of his report he gives some ash analyses but fails to interpret them. He also fails to give details as to the method of obtaining and estimating his ash. O'Brine's ash analyses are as follows:

KEY TO ASH ANALYSIS:
A = SiO_{2}.
B = Fe_{2}O_{3} and Al_{2}O_{3}.
C = CaO.
D = MgO.
E = K_{2}O.
F = Na_{2}O.
G = H_{2}SO_{4}
H = Cl.
I = P_{2}O_{5}.
J = CO_{2}

------+-----+-----+-----+-----+----+-----+----+----+----+----+-----+
Plant.|Total| | | | | | | | | | |
|ash. | A | B | C | D | E | F | G | H | I | J |
------+-----+-----+-----+-----+----+-----+----+----+----+----+-----+
AM |12.15|32.77|16.26| 6.05|3.11|13.30|3.21|3.9 |0.47|6.12|10.55|
| | | | | | | | | | | |
AL |13.52|17.08|12.21|14.27|2.62|17.26|5.75|3.22|3.87|3.30|17.37|
| | | | | | | | | | | |
AC |12.36| 7.82| 5.97|12.10|3.55|23.35|3.38|5.56|9.0 |4.67|20.62|
| | | | | | | | | | | |
------+-----+-----+-----+-----+----+-----+----+----+----+----+-----+
KEY TO PLANTS:
AM = _Astragalus mollissimus_ (whole plant)
AL = _Aragallus lamberti_ (whole plant)
AS = _Astragalus caryocarpus_

These analyses are evidently incorrect, as O'Brine estimates a carbon content of 4.13 per cent for the first, and for the second 2.22 per cent, showing incomplete combustion.

Mayo[133] experimented with alcoholic and aqueous extracts of dried _Astragalus mollissimus_ on guinea pigs, with negative results, and was first led to deny a relationship between the disease and the plants. Later, as a result of the post-mortem findings, he was convinced that his first conclusion was wrong and that "the disease is certainly the result of animals feeding upon the loco-weed." Mayo says:

A careful survey of the experiments performed and observations
noted leads me to the opinion that the disease known as "loco" is
the result of malnutrition, or a gradual starvation, caused by the
animals eating the plants known as "loco weeds," either _Astragalus
mollissimus_ or _Aragallus lamberti_. If there is a narcotic
principle in the plant, chemists have failed to find it and a fluid
extract does not possess it, and a ton of the plant eaten by an
animal ought to contain enough of the poisonous properties to
destroy an animal.

Kobert[134] has also tested the activity of _Astragalus mollissimus_ and says, "Ich fand _Astragalus mollissimus_ ziemlich unwirksam."

Doctor McEackran[135] fed dried _Astragalus mollissimus_ and _Aragallus lamberti_ mixed with feed to a stabled animal for two months without result. (Animal not stated).[136] Similar negative experiments are reported from the State of Washington, but the amounts used were too small to form any conclusions.[137]

Mr. V. K. Chesnut[138] has busied himself with the loco problem, but mainly in an executive capacity, his own efforts being directed to the study of the relation of the loco plants to the disease on the range. He has done no laboratory work. Chesnut and Wilcox made numerous autopsies on sheep and experiments on animals. They claimed that an extract of _Aragallus spicatus_ produced some slight narcotic action in rabbits. Their pathological examinations failed to show any characteristic lesion, but they state that the cerebral membranes were in all cases slightly congested. They deny any causative relationship to the presence of worms or with feeding upon alkalis. They believe that sheep are more likely to become locoed if not salted regularly. Chesnut describes one case in which a lamb became locoed by nursing from a locoed mother.

In 1901 Reid Hunt, at that time a special agent of the United States Department of Agriculture, studied the loco question in Montana, working mainly with _Aragallus spicatus_. He moistened the ground-up plant with 93 per cent ethyl alcohol and then percolated it until exhausted. This extract was evaporated and taken up with water so that 1 c.c. of the solution corresponded to 10 grams of the plant. This was fed to an active young rabbit weighing 490 grams, 6 c.c. being fed by the mouth and followed in about an hour by 10 c.c. more, and two hours after this by 15 c.c. This rabbit showed no symptoms during the following day. The next day it was very dull and there was marked muscular weakness, as the rabbit's legs were spread wide apart and his nose rested on the ground. Later respiration became very slow and the pupils were dilated. The paralytic symptoms increased and finally, after a convulsive movement, the animal died, thirty-six hours after the first feeding. Hunt merely states of the post-mortem examination that the stomach was well filled and that the "walls seem normal."

Hunt tried to isolate an active principle by the Dragendorff method, but failed to obtain any physiologically active shakings. He tried hypodermic injections of 80 per cent alcohol extractions of the fresh green plant, and after the injection of an extract corresponding to 60 grams of the fresh plant there was no effect produced. He tried to induce symptoms by feeding the plant itself to rabbits, but was unsuccessful, as the rabbits refused to eat the plant. He was not able to induce symptoms with the extracts of the dried plant.[139]

Marshall[140] studied the loco question with regard to sheep and practically denies the existence of a locoed condition due to eating the loco plants, but believes the condition due to bad feeding, parasitism, etc. He lays great stress upon the presence of worms, but fails to see that they may be merely a secondary infection superimposed upon an already morbid condition produced by eating the plants. Others have claimed that the cause is an insect living upon the loco plants. Others, again, have suggested an analogy with trypanosome disorders.

Chesnut has held the view that many of the cases of so-called locoed sheep were really due to parasites, but that there was a true locoed condition due to eating the loco weeds.

The lack of agreement in the results of the investigators has caused many to doubt any positive relation between the plant and the disease, and even as late as 1904 Payne[141] practically says these diseases are due to lack of nutrition and not to the loco plant. The matter has been summed up in a recent work as follows:

Though many chemists have sought for the constituents, none have
been able to locate the active properties, the trace of alkaloids,
resins, volatile and fixed oils having each in turn been found
destitute of it. Yet the poisonous properties are fully established
by field observations. The destructiveness of these plants to stock
is so great as to have probably caused upward of a million dollars
loss in the aggregate, and large bounties have been offered by
State governments for an effective method of avoiding such losses.
It is considered very probable that the poisonous constituent is
albuminoidal.[142]

* * * * *

FOOTNOTES:

[83] Storke, B. F. The Loco Weed. Med. Current, vol. 8, p.
155. 1892.--Kellogg, A. California and Colorado "Loco"
Poisons. Cal. Acad. Sci. Proc. for 1875, vol. 6, p. 3. 1876.

NOTE.--The very early reports of these loco plants were
purely botanical. See Torrey, J., Botany, in Report on the
United States and Mexican Boundary Survey, by W. H. Emory,
vol. 2, p. 56, 1859; also Botanical Register, London, vol.
13, pl. 1054, 1827.

[84] Vasey, G. Plants Poisonous to Cattle in California.
Rept. of Commissioner of Agriculture for 1874, p. 159. 1875.

[85] Vasey, G. Botanical Notes. Monthly Reports of Dept.
Agriculture for 1873, p. 503. 1874.

[86] Vasey, G. Botanical Notes. Monthly Reports of Dept.
Agriculture for 1874, p. 513. 1875.

[87] Brewer, W. H., and Watson, S. Geological Survey of
California, Botany, vol. 1, p. 155. 1876.

[88] Rothrock, J. T. Notes on Economic Botany, in G. M.
Wheeler's Report upon U. S. Geographical Surveys West of the
One Hundredth Meridian, vol. 6, p. 43. 1878.

[89] Kellogg, A. California and Colorado Loco Poisons. Cal.
Academy of Sciences, Proc., 1875, vol. 6, p. 3. 1876.

[90] Rothrock, J. T. Poisonous Properties of the Leguminosæ.
Acad. of Nat. Sci., Phila., Proc., vol. 29, p. 274. 1877.

[91] Prescott, A. B. Laboratory Notes--A Partial Analysis of
the Oxytropis Lamberti. Amer. Journ. Pharm., vol. 50, p. 564.
1878.

[92] Rept. of Commissioner of Agriculture for 1878, p. 134.
1879.

[93] Rept. of Commissioner of Agriculture for 1879, pp. 89,
90. 1880.

[94] Rept. of Commissioner of Agriculture for 1886, p. 75.
1887. Rept. of Commissioner of Agriculture for 1884, p. 123.
1884.

[95] Rothrock, J. T. Notes on Economic Botany, in G. M.
Wheeler's Report upon U. S. Geographical Surveys West of the
One Hundredth Meridian, vol. 6, p. 43. 1878.

[96] Gibbons, H. Poisonous Effects of Crotalaria--Vulgo
Rattle Weed, Loco Weed. Pacific Med. and Surg. Journ., vol.
21, p. 496. 1878-79.

[97] Ott, I. Physiological Action of Astragalus Mollissimus.
New Remedies, vol. 11, p. 227. 1882.

[98] Hill, J. R. Note on a Species of Astragalus from Cyprus.
Pharm. Journ. and Trans., 3 s., vol. 18, p. 712. 1887-88.

[99] Sayre, L. E. Loco-Weed. Proc. Amer. Pharm. Assoc., vol.
36, p. 112. 1888.

[100] Sayre, L. E. Loco Weed. Amer. Vet. Rev., vol. 11, p.
556. 1887.

[101] Kennedy, J. Loco Weed (Crazy Weed). Pharm. Rec., vol.
8, p. 197. 1888.

[102] Day, M. G. Experimental Demonstrations of the Toxicity
of the "Loco Weed." N. Y. Med. Journ., vol. 49, p. 237. 1889.

[103] Presumably a 10 per cent decoction, U. S. P.

[104] Storke, R. F. The Loco Weed. Med. Current, vol. 8, p.
157. 1892.

[105] Day, M. G. The Separation of the Poison of the "Loco
Weed." N. Y. Med. Journ., vol. 50, p. 604. 1889.

[106] Sayre, L. E. Active Principle of Loco Weed. Notes on
New Remedies, vol. 2, No. 12, p. 1.

[107] Day, M. G. Loco Weed, in F. P. Foster's Reference-Book
of Practical Therapeutics, vol. 1, p. 588. 1896.

[108] Stalker, M. 1st Ann. Rept. State Vet. Surg. Iowa, p.
16. 1885.

[109] Power, F. B., and Cambier, J. Chemical Examination of
Some Loco-Weeds. Pharm. Rundschau, vol. 9, p. 8.
1891.--Power, F. B. Notes on the So-called Loco Weeds. Pharm.
Rundschau, vol. 7, p. 134, 1889.--See also Hoffmann, F.,
Loco-Weeds, in Pharm. Rundschau, vol. 7, p. 168. 1889.

[110] Kennedy, J. Pharm. Rec., vol. 8, p. 197. 1888. Kennedy
also obtained ammonia from _Astragalus mollissimus_.

[111] O'Brine, D. Progress Bulletin on the Loco and Larkspur.
Colo. State Agric. Coll. Bul. 25, p. 18. 1893.

[112] Oatman, H. C. The Poisonous Principle of Loco Weed.
Notes on New Remedies, vol. 4, p. 14. 1891-92.

[113] Sayre, L. E. Loco Weed. Kans. Acad. Sci. Trans., vol.
18, p. 141. 1903.

[114] Sayre, L. E. Loco Weeds. 7th Bienn. Rept. Kans. State
Board Agric. for 1889-90, vol. 12, pt. 2, p. 99. 1891.

[115] Sayre, L. E. Further Report on Loco Weeds. Notes on New
Remedies, vol. 4, p. 80. 1891-92.

[116] Sayre, L. E. The Loco Disease. Journ. Kans. Med. Soc.,
vol. 4, pp. 241-243. 1904.--What is Insanity in Lower
Animals? Journ. Kans. Med. Soc., vol. 4, p. 222. 1904.

[117] Sayre, L. E. Loco Weed. Kans. Acad. Sci. Trans., vol.
18, p. 144. 1903.

[118] Ruedi, C. Loco Weed (Astragalus Mollissimus): A
Toxico-Chemical Study. Trans. Colo. State Med. Soc., p. 418.
1895.--Also Treatment of Animals Poisoned by Loco Weed
(unpublished article).

[119] The "Loco Disease." Therap. Gaz., vol. 12, p. 30. 1888.

[120] Sayre, L. E. Loco Weed. Kans. Acad. Sci. Trans., vol.
18, p. 142. 1903.

[121] Sayre, L. E. Loco Weeds. 7th Bien. Rept. Kansas State
Board Agric. for 1889-1890, pt. 2, p. 98. 1891.

[122] Curtice, C. Tape-Worm Disease of Sheep of the Western
Plains. Bur. Animal Industry, 4th and 5th Ann. Rept., p. 167.
1889.

[123] Steele, C. D. New Theory about Loco. Farm and Ranch,
vol. 20, No. 35, p. 1. 1901.

[124] Marshall, H. T. Loco Weed Disease of Sheep. Johns
Hopkins Hospital Bul., vol. 15, p. 181. 1904.--Data as to
these parasites of sheep may be found in Curtice, C., The
Animal Parasites of Sheep, Bur. Animal Industry, Rept., 1890.

[125] Faust, E. S., and Tallquist, T. W. Ueber d. Ursachen
der Bothriocephalus-anämie. Arch. f. Exp. Path., vol. 57, p.
367. 1907.

[126] Walshia Amorphella and the Loco Weed. Insect Life, vol.
2, p. 50. 1889-90. Snow, F. H. Loco-Weed. Science, vol. 9, p.
92. 1887.

[127] Lloyd, J. U. Loco, or Crazy Weed. Eclectic Med. Journ.,
vol. 53, p. 482. 1893.

[128] Lloyd, J. U., l. c., p. 483.

[129] Lloyd, J. U., l. c., p. 484.

NOTE.--Eccles had previously announced a somewhat similar
idea. Sayre, L. E. Loco Weed. Proc. Amer. Pharm. Assoc., vol.
36, p. 115. 1889.

[130] Lloyd, J. U., l. c., p. 486.

[131] Lloyd, J. U., l. c., p. 483.

[132] O'Brine, D. Progress Bulletin on the Loco and Larkspur.
Colo. State Agric. Coll. Bul. 25, p. 17. 1893.

[133] Mayo, N. S. Some Observations on Loco. Kans. State
Agric. Coll. Bul. 35, p. 116. 1893.

[134] Kobert, R. Lehrb. d. Intoxikationen, p. 615. 1893.

[135] O'Brine, D. Progress Bulletin on Loco and Larkspur.
Colo. State Agric. Coll. Bul. 25, p. 13. 1893.

[136] After the manuscript of this bulletin was sent to the
printer it was learned through Professor Carpenter that the
animal was a horse.

[137] Nelson, S. B. Feeding Wild Plants to Sheep. Bur. Animal
Industry, Bul. 22, p. 12. 1898.

[138] Chesnut, V. K., and Wilcox, E. V. Stock-Poisoning
Plants of Montana. U. S. Dept. Agric., Div. Bot., Bul. 26, p.
95. 1901.--Wilcox, E. V. Plant Poisoning of Stock in Montana.
Bur. Animal Industry, 17th Ann. Rept., p. 111. 1900.

NOTE.--The writer wishes to acknowledge the great literary
help Mr. Chesnut's card catalogue has been to him in the
preparation of this paper.

[139] Unpublished report.

[140] Marshall, H. T. Loco Weed Disease of Sheep. Johns
Hopkins Hospital Bul., vol. 15, p. 182. 1904.

[141] Payne, J. E. Cattle Raising on the Plains. Colo. Agric.
Expt. Sta. Bul. 87, p. 16. 1904.

[142] National Standard Dispensatory, p. 868. 1905.

NOTE.--The field experiments of Harding and Tudor are rather
conclusive as to the relation of these plants to this
disorder. Sayre, L. E., Loco Weed, Amer. Vet. Rev., vol. 11,
pp. 553-554, 1887--Blankinship, J. W., Loco and Some Other
Poisonous Plants in Montana, Mont. Agric. Exper. Sta. Bul.
45, pp. 83-84, 1903--Loco Disease, Therap. Gaz., vol. 12, p.
30. 1898.

=NOTES ON VARIOUS MEMBERS OF THE LOCO-WEED FAMILY.=

_Astragalus caryocarpus_ is at times eaten in some of the Western States, but is claimed by some at certain stages of its growth to contain a poisonous principle. Frankforter,[143] from experiments on himself, however, denies this.

_Astragalus glycophyllus_ has been used as a diuretic and _Astragalus exscapus_ in the treatment of syphilis.[144] "The seed of _A. boeticus_, planted in Germany and England, are found to be the very best substitute for coffee yet tried, and so used--roasted, parched, and mixed with coffee."[145] _Astragalus nuttallianus_, according to Smith,[146] is a highly nutritious forage plant in spring. _Astragalus crassicarpus_ has been prophesied by him to be a valuable addition to early spring soiling crops. _Astragalus adsurgens_ (_nitidus_) and one or two other species of Astragalus are still used in Chinese medicine.[147] The Indians of the Southwest are familiar with certain loco plants.[148] The Tewans of Hano are said to eat the root of _Aragallus lamberti_, and _Astragalus mollissimus_ is applied locally for headaches by some of the Arizona Indians. One of these species is used as a flavoring material by the Coahuillas and is mixed with other plants as spices.[149] _Astragalus kentrophyta_ had a reputation among the Navajos for the treatment of rabies.[150] The use of certain loco plants--_Astragalus mollissimus_--has been advocated on theoretical grounds in the treatment of certain forms of insanity, but without favorable results.[151] In Peru and Chile _Astragalus garbancillo_, _A. unifultus_, and _A. ochroleucus_ have been considered injurious to animals.[152] _Astragalus glyciphyllus_ and _A. alpinus_ have been used in Europe as food for stock.[153]

Details as to the use of other Astragali can be found in Planchon, G., Sur les Astragales, in Journal de Pharmacie et de Chimie, 5th series, vol 24, p. 473, 1891; 5th series, vol. 25, pp. 169, 233, 1892.

* * * * *

FOOTNOTES:

[143] Frankforter, G. B. A Chemical Study of Astragalus
Caryocarpus. Amer. Journ. Pharm., vol. 72, p. 320. 1900.

[144] Maisch, J. M. Poisonous Species of Astragalus. Amer.
Journ. Pharm., vol. 51, p. 240. 1879.--Fleurot. Chimiques et
Pharmaceutiques sur la Racines d'Astragale sans Tiges. Journ.
de Chim. Med., vol. 10, p. 656. 1834.

[145] Porcher, F. P. Resources of the Southern Fields and
Forests, p. 204. 1869.

[146] Smith, J. G. Fodder and Forage Plants. U. S. Dept.
Agric., Div. Agrost., Bul. 2 (rev. ed.), p. 12. 1900.

[147] Holmes, E. M. Notes on Chinese Drugs. Pharm. Journ. and
Trans., vol. 21, 3 s., p. 1149. 1891.

[148] Hough, W. Environmental Interrelations in Arizona.
Amer. Anthropologist, vol. 11, pp. 143, 147. 1898.

[149] Barrows, D. P. Ethno-Botany of the Coahuilla Indians of
Southern California, p. 67. 1900.

[150] Matthews, W. Navajo Names for Plants. Amer. Nat., vol.
20, p. 772. 1886.

[151] Givens, A. J. Loco or Crazy Weed. Med. Century, vol. 1.
p. 21. 1893.--Compare Hurd, H. M. Amer. Journ. Insanity, vol.
42, p. 178. 1885-86.

[152] Rosenthal, D. A. Synopsis Plantarum Diaphoricarum,
Erlangen, 1861, p. 1004. Greshoff, M. Beschrijving d. Giftige
en Bedwelmeude Planten bij de Vischvangst in Gebrulk, p. 51.
1900.

[153] Pott, E. Handb. d. tierisch. Ernährung, vol. 2, p. 113.
1907.

=LABORATORY EXPERIMENTS--PHYSIOLOGICAL.=

The first point in our investigations was to determine whether the plant exerted any poisonous action and to find some animal which responded regularly to it; then to ascertain if the lack of results of previous investigators was not due to insufficient doses, and later to see if by feeding smaller amounts at repeated intervals symptoms comparable to those described as occurring on the range could not be produced. The animal finally selected was the rabbit.

=EXPERIMENTS ON RABBITS.=

=ACUTE CASES.=

_Experiment No. 1._--On September 8, 1905, an aqueous extract of 333 grams of fresh _Astragalus mollissimus_, made in Hugo, Colo., and shipped preserved in chloroform,[154] killed a rabbit weighing 1,616 grams in one hour and thirty-five minutes, while an extract corresponding to 167 grams merely caused drowsiness and loss of appetite in a rabbit weighing 765 grams.

_Experiment No. 2._--On November 29, 1905, a rabbit weighing 1,162.3 grams was fed with a concentrated aqueous extract of 500 grams of fresh _Astragalus mollissimus_, which had been shipped from Hugo, Colo., preserved in chloroform in sealed vessels. This animal died in one hour and ten minutes. The symptoms consisted in dullness, rapid respiration, and signs of pain. At autopsy the stomach and upper part of the small intestines showed hemorrhagic ecchymoses, with dilation of the dural vessels of the brain and cord, with a clot over a portion of the spinal cord.

_Experiment No. 3._--On February 13, 1906, a rabbit weighing 992 grams was fed with a concentrated aqueous extract of 500 grams of the fresh _Astragalus mollissimus_, collected in September and preserved in chloroform water. Before feeding, the rabbit's ears were warm and the rabbit struggled when any attempt was made to turn him on his back. The temperature at 10.50 a.m., the time of feeding, was 103.5°F.; at 11.15 a.m., 102.5°F. At 11.30 a.m. the rabbit was breathing very rapidly and would stay on his back for some time if placed so. The temperature at this time was 102.6°F. Both pupils, the one exposed to the light and the one protected, were contracted. At 12.02 p.m. convulsive movements of the legs appeared. The rabbit made one leap, the temperature rose to 103.6°F., and after a few convulsive movements of the limbs the anus relaxed and a small stool appeared, the pupils dilated, and the animal died at 12.06 p.m.

_Experiment No. 4._--The feeding of the extract of 464 grams induced a fall in temperature of 2.4°F. in three hours, and the rabbit died several hours later (at night).

_Experiment No. 5._--March 2, 1906, a rabbit weighing 928 grams was fed with a concentrated extract of 500 grams of the fresh seeds and pods of _Astragalus mollissimus_, made in September, 1905, and preserved with chloroform water. This animal died in one hour and seven minutes. The animal showed the usual post-mortem conditions.

It was thus found that the aqueous extract of 500 grams of the fresh _Astragalus mollissimus_ would cause death in about one hour in rabbits weighing about 2 pounds (907 grams), these rabbits showing constant clinical symptoms--urination, paralysis, more or less convulsive muscular twitchings, often terminating in general convulsions, drowsiness, and stupor, with more or less anesthesia. The pupils at the time of death were often unequal. At first there was usually a slight rise in temperature, but this was soon succeeded by a fall. Often there were soft stools. The post-mortem lesions in these cases were marked congestion, with hemorrhages in the stomach walls and a secretion of thick mucus. The portions of the stomach walls most affected were the dependent portions near the cardiac end. The intestines showed dilatation of the blood vessels. The mesenteric vessels and also the vessels in the cerebral portions of the dura were markedly dilated; in some cases there were clots, especially at the posterior portion of the brain, between the cerebrum and the cerebellum. At times there were clots over the dorsal portion of the cord. On cutting into the brain the brain substance itself did not appear to be congested. The cord seemed about normal, but the vessels of its membranes were well marked. The other organs showed nothing characteristic macroscopically. These experiments were repeated many times and found to be constant.

These acute symptoms were likewise produced by an extract of 500 grams of the fresh _Aragallus lamberti_ from Arizona preserved in chloroform water (rabbit weighing 1,998 grams). An aqueous extract of 150 grams of the dried _Astragalus mollissimus_[155] from Imperial, Nebr. (1906), caused death in one hour and fifty-eight minutes in a rabbit weighing 1,530 grams, and an extract of 100 grams killed in one hour and twenty-two minutes a rabbit weighing 736 grams.

An aqueous extract of 100 grams of the dried _Astragalus bigelowii_ induced death in one hour and thirty-eight minutes, the rabbit weighing 1,502 grams.

An aqueous extract of 150 grams of _Astragalus nitidus_ collected at Woodland Park, Colo., in 1906 induced death in three hours and five minutes, the rabbit weighing 1,672 grams.

An aqueous extract of 200 grams of the dried _Astragalus bisulcatus_ caused death after several hours (at night), the rabbit weighing 2,423 grams.

In certain cases this production of acute symptoms was not entirely a question of salt action, as was shown by certain other experiments. In other cases salt action seems to be the important factor, so that the production of these acute symptoms can not always be considered characteristic.

* * * * *

FOOTNOTES:

[154] In all cases in which the plants were preserved with
chloroform sealed vessels were used for shipping. The
chloroform was carefully evaporated off in vacuo before
feeding the extract, the evaporation requiring several hours.
The plants were collected by Dr. C. Dwight Marsh, in charge
of the field investigations at Hugo, Colo.

[155] All extracts from dried material were made at
Washington.

=CHRONIC CASES.=

_Experiment No. 6._--February 19, 1906, a large gray rabbit weighing 2,055.3 grams was fed with 60 c.c. of fluid representing the concentrated aqueous extract of 250 grams of the fresh _Astragalus mollissimus_, collected September 18, 1905, and preserved in chloroform. This rabbit was very hard to hold. The ears rested on the body. The temperature at the time of feeding, 1.30 p.m., was 102.3°F. At 2.57 p. m. the animal looked dull but resisted handling. At 3.30 p.m. it urinated. At 4.15 p.m. the temperature was 98.5°F., the pupils were about the same size as before feeding, and the animal became much duller. The next day at 12.50 p.m. the temperature was 102.4°F., and at this time the animal could be handled with greater ease. The animal ate in the morning. The same amount of extract was again fed at 1.24 p. m. At 1.35 p.m. the animal was much duller and could be turned on his back with ease. If disturbed he ran against the wall as if utterly unconscious of the obstruction. The animal had soft, liquid, brown stools and tried to lie down as much as possible. If turned on its back with the feet up it would stay so almost indefinitely. Temperature, 103.8°F.; respiration very rapid. At 2.40 p.m. the temperature was 99.8°F., and the animal died a few minutes later. After death the pupils were much contracted. The vessels of the dura covering the brain were much dilated, but the vessels inside the brain were not dilated. The stomach walls were congested and marked with numerous petechiæ and covered with mucus.

_Experiment No. 7._--On February 19, 1906, a white and brown rabbit whose temperature was 103.2°F. was fed 30 c.c. of aqueous fluid representing the concentrated extract of 125 grams of the fresh _Astragalus mollissimus_, collected September, 1905, and preserved with chloroform. The rabbit weighed 1,502.5 grams. This extract was fed at 1.45 p.m., and at 4.15 p.m. the temperature was 102.6°F., but there were no marked symptoms. The following day at 2.04 p.m. the temperature registered 102.5°F. The same amount of extract was given at 2.09 p.m. The temperature at 4 p.m. was 99.8°F., the animal was dull, and the pupils were perhaps a little smaller. The animal could not be turned over without resistance. The following day, February 21, at 1.30 p.m. the temperature was 102.6°F., and at 1.45 the same amount of extract was given. At 1.54 p.m. the animal was much duller and the breathing was very rapid. At 4.10 p.m. the temperature was 101.3°F. The animal had been dull ever since the feeding was begun. It nibbled food shortly before the last feeding. On February 23 the same amount of extract was given at 2.16 p.m., temperature 99°F. The breathing was very rapid, the ears shaking, and there was a sleepy, dull look about the animal. At 3.30 p.m. the animal was dull, but would still walk about if disturbed. At this time the animal weighed 1,445.8 grams. At 4.30 p.m. the temperature was 102°F. and the pupils were about normal size. There was a marked sleepy look about the animal, which sat quietly in its cage.

February 24, at 1 p.m., the animal was very dull and could with ease be turned on its back with its feet in the air. It would sit in its cage perfectly quiet. The weight at this time was 1,417.5 grams, the temperature 96.6°F. On February 26 the animal weighed 1,360.8 grams. It was dull and refused to eat. The abdomen felt very distended and tympanitic. February 27 the weight was still 1,360.8 grams, and the animal sat in its cage as if asleep, with eyes half closed. There was no diarrhea and the abdomen was very distended. At 11.15 a.m. there was a general convulsion and the animal fell over. At 12 m. the abdomen seemed even more swollen, the animal was hardly able to walk, and it fell over, uttering a cry. Pupils were about normal--perhaps a little smaller. The animal died at 12.10 p.m.

The post-mortem, made immediately after death, showed the abdomen markedly tympanitic, and the large intestines could be outlined through the abdominal walls with ease. The large intestines were of a chocolate color, intensely congested, and marked with hemorrhages. On opening the abdomen there was a decided putrefactive odor, and about an ounce of bloody fluid was found in the peritoneal cavity, together with fibrin flakes. The stomach was pale, the first three inches of the small intestine up to where it turned sharply were pale, and below this the intestines were injected and full of gas and of a dark red color. The kidneys were 3-1/2 centimeters long and were pale, capsules easily peeled off; cortex pale. Liver pale and infected with some coccideæ. The gall bladder was one-quarter inch wide and one inch long. Spleen a trifle pale; lungs pale, nothing abnormal; heart relaxed. On opening the stomach gas and fluid, with some food, exuded. The walls were pale, but pink in some places. There was no marked congestion or hemorrhage or perforation. The mesenteric vessels were dilated. The upper portion of the intestines contained a little mucus-like fluid, but lower down became bloody, and still lower contained pus-like fluid. The walls were hemorrhagic. The large intestine contained a soft, fecal-like fluid, very foul. Its walls were much congested and full of hemorrhagic points. The cortex of the suprarenal bodies was sharply defined, the medullæ brownish. Brain pale, some dural vessels well marked, no clots or hemorrhages. Base of brain pale. No congestion seen on cutting into the brain. Spinal cord showed no hemorrhages or lymph effusions.

_Experiment No. 8._--On February 18, 1906, at 2 p.m., a rabbit whose temperature was 102.2°F. was fed with the aqueous extract of 125 grams of fresh _Astragalus mollissimus_, collected in September, 1905, and preserved in chloroform, 30 c.c. of the fluid being used. At 4.25 p.m. the temperature was 102.4°F. No symptoms were noted. This rabbit weighed 1,644.3 grams. On February 20 at 2.09 p.m. the temperature was 102.2°F. and the rabbit showed no symptoms. The same dose was repeated at 2.15 p.m. At 4 p.m. the temperature was 100.3°F. The rabbit was dull but could not be turned over without a struggle. February 21 at 1.30 p.m. the temperature was 101.4°F. The same amount of extract was fed at 1.45 p.m. At this time the animal was dull and breathed more rapidly. At 4.10 p.m. the temperature was 97.3°F. Next day the same amount of extract was again given at 2 p.m. At 2.16 p.m. the breathing became rapid and the animal duller. The ears were directed forward. At 4.15 p.m. the temperature was 101.6°F.; weight 1,757.7 grams; animal slightly dull. February 24, temperature 102°F., weight 1,786 grams. March 5, weight 1,729.3 grams. The animal was fed at 3.20 p.m. with a concentrated extract of 125 grams of _Astragalus mollissimus_, collected in September. Temperature at time of feeding 100.4°F.; 3.40 p.m., no symptoms; 4 p.m., temperature 102°F. March 7, weight 1,644.3 grams; March 8, weight 1,672.6 grams; March 10, weight 1,701 grams; March 12, weight 1,658.4 grams; March 14, weight 1,701 grams.

In this case, where the same dose was given in a period of five days, very little effect on the rabbit was noted.

_Experiment No. 9._--On March 1, 1906, a black rabbit weighing 2,664.8 grams was fed with a concentrated aqueous extract of 250 grams of fresh _Astragalus mollissimus_, collected in the fall of 1905.

On March 5 the weight was 2,296.3 grams. The animal was then given the same amount of extract. During the afternoon it passed mucus and thick pieces of feces and was dull; respiration very rapid. March 6, weight 2,282 grams; March 7, 3 p.m., animal very dull and would not eat; sat hunched up, but resisted being disturbed: weight 2,310.5 grams. March 8, weight 2,183 grams; March 9, weight 2,069.5 grams. Pupils dilated; finger could be run almost against the eye, provided the lashes were not touched, without the animal winking or paying any attention. Rabbit ate very little and had not urinated since the preceding day. Left ear had fallen to the side as if the animal were unable to support it. Weight, 1,912.8 grams. From March 9 to March 11, 67 c.c. of cloudy urine were voided. This did not clear with acetic acid. Left eye tearing. March 10, head held to right side. March 12, weight 1,786 grams. Left pupil smaller than right, neither responding to light. Rabbit very weak. March 14, weight 1,729.3 grams. Would not eat. March 16, weight 1,644.3 grams. Right pupil larger than left, neither responding to light. Diarrhea present. Breathing noisy. In sitting down she raised herself on her forelegs, evidently to take the pressure off her abdomen, which was distended. If disturbed, she would butt against the side of the cage, apparently oblivious of its presence. Knee jerks were very active, almost a clonus. Reflex from tendo Achillis active. March 17, forelegs spread out, head falling to left side. The temperature had fallen below 94°F. and would not register on the ordinary clinical thermometer. The ears twitched, the head was thrown back, the abdomen was distended, and the rabbit gritted its teeth. Died. Weight, 1,559.2 grams.

Brain and spinal cord pale. Dural vessels plainly seen but not marked. Intestinal vessels congested. Stomach pale; nothing apparent macroscopically save a small pin-point ulcer.[156] Heart relaxed. Post-mortem examination otherwise negative macroscopically.

_Experiment No. 10._--A mouse-colored rabbit weighing 1,927.8 grams was fed February 18, 1906, at 2.26 p.m., with a concentrated aqueous extract of 250 grams of fresh _Astragalus mollissimus_ collected in September, 1905, and preserved in chloroform water. The temperature of this rabbit was 102.6°F. The fluid given was 40 c.c. At 2.45 p.m. the rabbit urinated and at 2.57 p.m. was dull and the respiration became rapid. The animal then aborted and had three young, two of which showed some movement after birth, but were apparently premature.

On February 23 the temperature of this rabbit was 102.9°F. at 1.40 p. m. She was then fed with the same amount of the extract as before. At 2.16 p.m. she lay down and became much duller; left ear fallen to side. At 3.30 p.m. the rabbit was unable to stand. The pupil of the eye exposed to the light was dilated. The animal died without a struggle. The stomach contained much bloody mucus. In the dependent portion of the stomach near the cardiac end were marked petechiæ in the walls, with bright-red blood in the stomach itself. The heart was relaxed. The intestines showed nothing abnormal. The dural vessels of the brain were dilated; there was a clot on the dura over the fourth ventricle. Spinal cord and kidneys normal, the capsules not adhering. Weight, 1,786 grams at death.

_Experiment No. 11._--On March 1, 1906, a rabbit weighing 2,126.2 grams was fed with a concentrated aqueous extract of 250 grams of the fresh _Aragallus lamberti_ preserved in chloroform water. On March 5 this dose was repeated, 37.5 c.c. of the fluid being used. March 6 the rabbit weighed 1,956 grams; March 7, 1,913.6 grams; March 8, 1,828.5 grams; March 9, 1,701 grams; March 12, 1,672.6 grams; March 14, 1,644.3 grams.

_Experiment No. 12._ January 19, 1906, a concentrated aqueous extract of 500 grams of the fresh _Aragallus lamberti_ preserved with chloroform water was fed to a rabbit weighing 785 grams. The temperature at 12.10 p.m., the time of feeding, was 101.6°F. The temperature 1 hour and 43 minutes later was 94.6°F., and the animal died shortly after, showing the same condition as occurred after feeding extracts of _Astragalus mollissimus_.

* * * * *

FOOTNOTES:

[156] Compare Plönius, W., Beziehungen d. Geschwürs u. d.
Erosionen d. Magens z. d. funktionell. Störungen u. Krankh.
d. Darmes, Arch. f. Verdauungsk., vol. 13, pp. 180, 270,
1907, and Tixier, L., Anémies Exper. Conséc. aux Ulcér. du
Pylore, Comp. Rend. Hebd. Soc. de Biol., vol. 62, p. 1041,
1907.

=PREGNANT ANIMALS.=

_Experiment No. 13._--A large, gray, pregnant rabbit weighing 2,891.6 grams was fed on February 22, 1906, with 42 c.c. of fluid, corresponding to the aqueous extract of 250 grams of _Astragalus mollissimus_ collected in September and October, 1905, and preserved with chloroform. At 4 p.m. the animal was dull, but still resisted efforts to handle. On February 24 this animal weighed 2,778.2 grams, and on February 26 it bore a litter of seven young rabbits. One or two of these showed movements of the limbs, but were apparently immature. This rabbit on March 10 weighed 2,537.3 grams; March 12, 2,438 grams; March 14, 2,508.9 grams; March 22, 2,494.7 grams.