LABORATORY EXPERIMENTS—CHEMICAL.

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.

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.

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 ofAstragalus mollissimuscollected 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.

Experiment No. 14.—On March 1, 1906, a black rabbit weighing 2,721.6 grams was fed at 12.15 p.m. with a concentrated aqueous extract of 250 grams of the freshAstragalus mollissimuscollected in September, 1905. On March 2 it weighed 2,438 grams; at 2.58 p.m. it still resisted efforts to turn it on its back; at 3.15 p.m. it could be turned on its back with ease. March 6 the weight was 2,338.8 grams;March 7 the animal was very dull, would not eat, pupils dilated, hind legs paralyzed; died during the night; weight, 2,267.9 grams.

The stomach walls were pale save at the dependent portion near the cardiac end, where there was a hemorrhagic, ulcerated area about 1-1/2 by 1-1/2 inches. The intestines were full of gas, but not hemorrhagic. The uterus contained eight immature fœti. The uterine walls were hemorrhagic. The kidneys weighed 9-1/2 grams; their medullæ were dark and the straight tubules well defined. The cerebral dural vessels were congested and the spinal dural vessels were well defined. The bladder was found contracted. The blood gave no bands for methæmoglobin, but showed merely those of oxyhæmoglobin on spectroscopic examination.

Experiment No. 15.—Control experiments made by feeding water were negative, except when a large quantity (150 c.c.) of water was given to a rabbit weighing 1,020.5 grams. The animal died in 12 hours with marked pallor of the tissues (hydræmia), a pathological condition quite different from that obtained by feeding extracts of the loco plants, and no such results were secured with the amount of water used in our feeding experiments, 50 to 70 c.c.

Experiment No. 16.—On February 28, 1906, a white rabbit weighing 581.2 grams was injected subcutaneously at 10.35 a.m. with a concentrated aqueous extract of 83 grams of freshAstragalus mollissimuscollected in September, 1905, and preserved with chloroform. The temperature before injection was 102.1°F. At 1.40 p.m. the animal was dull; at 3.12 p.m. the temperature registered 99.8°F. The animal died during the night. The post-mortem examination was negative. Stomach pale; heart relaxed save left ventricle, which seemed contracted; dural vessels of the brain dilated; kidneys perhaps normal. No microscopical examination.

Experiment No. 17.—February 28, 1906, at 10.25 a.m., a guinea pig weighing 496 grams was injected subcutaneously with a concentrated aqueous extract of 83 grams of the freshAstragalus mollissimuspreserved in chloroform water. At 1.40 p.m. there was muscular twitching. The animal was dull and could be easily turned on his back. The hind legs began to show weakness. At 1.50 p.m. the hind legs were almost completely paralyzed and the animal could be easily turned on his back. Muscles of the limbs twitched and semen was expelled. Animal died at 2.15 p.m.

Post-mortem showed dural vessels of cord and brain full of blood. Stomach pinker than normal: mesenteric vessels dilated. Heart almost empty of blood. Kidneys congested.

These experiments indicate that an acute form of poisoning may be induced by feeding concentrated aqueous extracts ofAstragalus mollissimusandAragallus lambertifrom Hugo, Colo., and Imperial, Nebr., to rabbits, and that if the extract is given in smaller and repeated doses a more prolonged or chronic condition may follow.

The rabbits showing the chronic effects of these plants exhibit symptoms which have a marked parallelism with those reported as occurring in larger herbivora (horses and cattle) on the range when locoed; that is, the loss of appetite (Experiment No. 9), the emaciation and loss in weight (Experiment No. 9), the dullness and stupor, with more or less anesthesia (Experiment No. 7), the disturbance in the visual function (Experiment No. 9), and the mental symptoms (Experiment No. 6). The occasional abortion compares with what has been observed in larger animals. The driedAstragalus mollissimusandAragallus lambertistill retained their poisonous properties, as we were able to kill with aqueous extracts of the dried plants made in the laboratory under the proper conditions.

Experiment No. 1.—On May 31, 1906, a sheep weighing 32.2 kilos was fed with a concentrated aqueous extract of 1,000 grams of the freshAstragalus mollissimuspreserved in chloroform water. The temperature at 11 o’clock, the time of feeding, was 103.4°F. At 11.45 a.m. this dose was repeated. At 12 o’clock the temperature was 104.1°F. At 12.45 the animal urinated. At 1.10 p.m. a similar extract of 2,000 grams was fed. The total liquid used was 1,500 c.c. On June 1 no symptoms were noted. On June 5 an extract of 3,000 grams of freshAragallus lambertiand 3,000 grams ofAstragalus mollissimuswas fed. After feeding this the animal could be easily turned over on its back and its ear pricked with impunity. The animal at this time weighed 30.8 kilos. On June 6, at 11 a.m., the temperature was 104°F. The sheep had numerous soft stools, and was very dull, and would not eat. On June 7 the temperature was 103.7°F. and the sheep still refused to eat. On the 8th the temperature was 103.2°F. at 10.40 a.m., and the stools were still numerous and soft.

There were then fed 640 c.c., representing the aqueous extract of 4,000 grams of the freshAragallus lamberti. The animal could be easily turned on its back. It weighed at this time 28.57 kilos. On June 9, at 10.47 a.m., the temperature was 103.4°F. The sheep still did not eat, but had no diarrhea. It now weighed 27.9 kilos, and the temperature was 103°F. at 10.45 a.m.

On June 13 the animal began to eat, and 1,700 c.c. of fluid, representing 5,500 grams of the freshAragallus lamberti, were fed. The temperature at 12.30 p.m. was 103°F. On June 14 the temperature was 103.4°F., the animal weighed 28.3 kilos, and refused food. On June 16 the weight was 28.3 kilos; the temperature at 2 p.m. was 103.5°F. There was no diarrhea.

On June 19 the aqueous extract of 1,000 grams of the driedAstragalus mollissimuswas fed with 420 c.c. of water. The temperature was 102.6°F. On June 20 the temperature was 102.9°F. at 10.45 a.m.

On June 21 500 c.c., representing the aqueous extract of 1,000 grams of the driedAstragalus mollissimus, were again fed. The animal now weighed 26.9 kilos. On June 26 the animal weighed 26 kilos, and its gait was very uncertain. The temperature was 104.2°F. It was fed 300 c.c. of fluid, representing the extract of 400 grams of the driedAstragalus mollissimus. On June 29 the animal weighed 26.8 kilos and the temperature was 102.8°F. It was fed the extract of 1,000 grams of driedAstragalus mollissimusin 500 c.c. of water. On June 30, at 10.45 a.m., the temperature was 104.2°F. The animal was very dull and died at night.

At autopsy the intestines and stomach merely appeared pale. There were no worms, and the lungs and other organs appeared normal.

Experiment No. 2.—A lamb weighing 15.4 kilos was fed on July 6, at 1.10 p.m., with 640 c.c. of fluid, representing the extract of 2,000 grams ofAstragalus mollissimus. At 1.17 p.m. the animal could be turned on its back, and it regained its feet with difficulty. At 1.24 p.m. it urinated and had a stool. The lamb died during the night.

The autopsy the following morning showed the heart filled with clots; lungs normal save for hypostatic congestion. The cerebral and dural vessels were dilated. About 1-1/2 teaspoonfuls of bloody serum were found at the base of the brain. There was none in the lateral ventricles, and no clots. The kidneys exhibited no marked congestion. There was no fluid found in the peritoneal or the pleural or pericardial cavities. The first stomach, however, contained small hemorrhagic spots, and the second was black. There were small hemorrhages in the intestines.

Experiment No. 3.—July 13, 1906, a sheep weighing 19.5 kilos was fed with 640 c.c. of fluid, representing the extract of 2,000 grams ofAragallus lamberti. The temperature at the time of feeding, 1.10 p.m., was 105.3°F. At 1.49 p.m. the sheep could be easily turned on its back. At 2.23 p.m. the temperature was 103.6°F. At 3.42 p.m. the temperature was 103.5°F. At 4.20 p.m. the respiration was fairly rapid. On July 14, at 11.15 a.m., the temperature was 103.6°F. The sheep would run about but could easily be turned over. It had not eaten, but there was diarrhea present. July 15, at 3.30 p.m., the temperature was 104°F. The animal had eaten. On July 17 the temperature was 104°F. and the animal weighed 18.8 kilos. On the 27th it weighed 17.2 kilos; on August 29, 20.8 kilos.

Experiment No. 4.—A lamb weighing 19 kilos was fed August 21, 1906, with 740 c.c., representing the aqueous extract of 2,500 grams of the freshAstragalus mollissimus, shipped to Washington in September, 1905. This animal ate at night, but the following day was dull. When seen on August 27 there was diarrhea present and the animal was still dull. On the 28th the animal died, weighing 16.7 kilos. There was no autopsy on account of decomposition.

Experiment No. 5.—A lamb weighing 15.6 kilos was fed on September 4, 1906, with an aqueous extract representing 3,500 grams of the driedAragallus lamberti, 1,000 c.c. of water being used. The temperature at the time of feeding was 104.3°F. At 2.48 p.m. the animal on rising to its feet developed a slight tremor of the fore legs and showed marked disinclination to stand on its feet. The temperature was 104°F. The animal died at 4.25 p.m. The post-mortem was negative, save for some reddening of the second stomach.[157]

These feeding experiments in sheep can not be considered quantitative, because, as is shown later, aqueous extracts of dried plants are often inactive, yet poisonous principles may be obtained from the plants by treatment with digestive fluids.

Extracts of dried loco plants vary much in their toxicity; with some the writer was unable to kill rabbits, even when an extract of 300 grams of the dried plant was used. It is interesting to note that when the field station was established at Hugo, Colo., in 1905, almost all the aqueous extracts of dried specimens sent to Washington would produce the acute symptoms of poisoning in rabbits, but during the third season of its existence many of the samples sent from the same area were much less active, if not inactive.

FOOTNOTES:[157]There was a slight odor of chloroform noticed on opening the stomach, so that perhaps the imperfect removal of the chloroform due to a hurried evaporation of the extract should be taken into consideration in this case.

FOOTNOTES:

[157]There was a slight odor of chloroform noticed on opening the stomach, so that perhaps the imperfect removal of the chloroform due to a hurried evaporation of the extract should be taken into consideration in this case.

The fact that the aqueous extract of 500 grams of the freshAstragalus mollissimus, or of 200 grams (in some cases 100 grams) of the dried plant, when fed by mouth, would regularly kill a rabbit weighing about 907 grams, with certain definite clinical symptoms and pathological lesions, was at first arbitrarily selected as our testto aid in the isolation of the active principle. Later the production of chronic symptoms by the aqueous extract or digestion of 200 grams of these dried plants given in doses of 100 grams each on two successive days was considered essential. Carnivora, such as dogs and cats, vomit so easily as to render them unsuitable for these investigations. The aqueous extract was distilled with and without steam, also after acidifying with sulphuric acid, and likewise after the addition of magnesium oxid, but in all cases the distillate was inactive.

The concentrated aqueous extract was shaken by the Dragendorff method with petroleum ether, benzol, chloroform, ether, and amyl alcohol, both in alkaline and acid condition, but the shakings yielded no physiologically active body. Shakings by the Otto-Stas method also proved inactive. Lead acetate, lead subacetate, silver nitrate, mercuric chlorid, alcohol, phosphotungstic acid, trichloracetic acid, ammonium hydrate, sodium carbonate, sodium hydrate, Mayer’s solution, uranyl acetate, silver oxid, and barium carbonate also failed to remove the active constituent. They gave heavy precipitates in all cases, but these proved inactive. Hydrocyanic acid was sought for with negative results. The pathological lesions in the very acute cases suggested in some respects oxalic acid, a saponin, a metal, or perhaps a toxalbumin as the active principle, but none of the precipitants for saponins, such as lead and copper, or the magnesium oxid method yielded a body which was active. Proteids were excluded by the fact that the various proteid precipitants—alcohol, trichloracetic acid, lead subacetate, mercuric sulphate or chlorid, and salting out with ammonium sulphate and sodium chlorid (complete saturation and half saturation)—failed to give an active precipitate. Glucosidal or alkaloidal bodies were also excluded. On dialysing for twenty-four hours, some of the active principle went into the dialysate and some remained in the dialyser. Ether yielded a precipitate from alcoholic solution which failed to kill. The possibility of the activity of the plants being due to its normal acidity was excluded by neutralizing the extract with sodium hydrate and precipitating the salts with alcohol. The filtrate proved active after removing the alcohol.

The negative results in looking for active alkaloidal, or glucosidal, or proteid bodies suggested that perhaps the action was due to some inorganic constituent. The writer then boiled the extract three minutes and as the filtrate was still found active and the proteid precipitate inactive became convinced of the inorganic nature of the active constituents, and finally incinerated the plant. The acid extract from this was also active, but death was delayed several hours. This was believed to be due to the insoluble form into whichthe compound was converted.[158]In fact, the question of solubility and the avoidance of an acid reaction, which of itself may kill, are the main points to keep in mind.

These experiments indicated that the injurious action toward rabbits of theAstragalus mollissimusandAragallus lamberticollected at Hugo, Colo., was due to one or more inorganic constituents,[159]but it does not follow that all loco plants have the same poisonous principle nor that the same species occurring on all soils has the same poisonous action.[160]

OfAstragalus mollissimusfrom Imperial, Nebr., collected in 1906, 200 grams were ashed in a platinum bowl and extracted with water. This aqueous extract when neutralized produced no marked symptoms in a rabbit and the weight of the animal remained about the same.

The ash undissolved after this extraction was then treated with acetic acid and water overnight, and after carefully evaporating off the acetic acid on the bath (tested by litmus paper) the residue was fed, partly in solution and partly suspended in water, to a rabbit weighing 1,800.2 grams. Next day the rabbit weighed 1,771.8 grams, showed paralysis of the limbs, and died during the morning. The stomach was intensely reddened and contracted.

An extract of a similar ash was made by boiling the same amount with a large quantity of 94 per cent alcohol. This was evaporated in vacuo and taken in water and fed to a rabbit weighing 1,459.9 grams. On the sixth day the animal died, having lost 70.9 grams in weight. The stomach showed reddening but no ulcers.

An acetic acid aqueous extract, made from the ash after the alcoholic extraction, proved inactive, showing that the alcohol had removed the active bodies. A 70 per cent alcohol extract of another ashed lot proved active, killing the rabbit overnight.

OfAstragalus mollissimusfrom Imperial, Nebr., 200 grams were ashed in a platinum bowl and the ash treated with acetic acid water. After freeing from acid, one half of the solution and emulsion was fed one day and the second half fed the following day. The rabbit at the time of feeding weighed 1,275.7 grams. Fourteen days later the animal died, weighing 1,105.6 grams. No autopsy.

A similar extract of the ash from between 100 and 150 grams of the same dried plant produced death in a rabbit weighing 1,190 grams in two hours and fifty-eight minutes.

The acetic acid extract of the ash of 125 grams of a mixture of the driedAstragalus mollissimusandAragallus lambertireceived from Hugo, Colo., June, 1907, after freeing from acid, was fed to a rabbit weighing 1,304 grams on July 29. On July 30 it weighed 1,332.4 grams. August 1 it weighed 1,219 grams, and it died the same day. The stomach was reddened and showed ulcers.

A similar extract from 250 grams of the same dried plants on boiling gave a heavy precipitate, but this precipitate was inactive, while the filtrate killed a rabbit in four hours.

Of dryAragallus lamberticollected in September, 1906, 200 grams were extracted with water and fed to a rabbit weighing 1,516.7 grams. Two days later the animal weighed 1,360 grams and died the same day.

The ash from 200 grams of the same dried plant was extracted with acetic acid, and after evaporating off the acid this was fed to a rabbit weighing 2,045.3 grams. Seven days later the animal weighed 1,729.3 grams, having lost 316 grams in weight.

The ash from 250 grams of the same species of plant, after similar treatment with acetic acid, induced death in a rabbit weighing 2,069 grams in 2 hours and 20 minutes. The stomach was inflamed.

FOOTNOTES:[158]Work is now being done by the writer on the inorganic constituents of various plants.[159]Scattered throughout the veterinary literature one finds cases of poisoning in animals with symptoms similar to those occurring in locoed animals which are attributed to eating plants grown on a peculiar soil, as in Oserow, Ueber Krankh. d. Pferde, welche Aehnlichkeit mit der Cerebro-spinal meningitis haben, aber durch Vergiftungen mit Gräsern von Salzgründen (Salzmooren) verursacht werden, Journ. f. Allgem. Veterinär-Medicin, St. Petersburg, p. 486, 1906. Abstract in Jahresber. über d. Leistungen auf dem Gebiete d. Veterinär-Medicin, vol. 26, p. 226, 1906.—Compare also Étude sur Quelques Plantes Vénéneuses des Regions Calcaires, Bul. Soc. Cent. de Méd. Vét., vol. 48, p. 378. 1894.[160]After completing this work the writer found that Sayre had said that he "had the suggestion that the harm coming from this plant is due to the inorganic constituents; this clue has been followed up, but like the others has brought us no nearer to the solution of the problem." Kans. Acad. Sci. Trans., vol. 18, p. 144. 1903.

FOOTNOTES:

[158]Work is now being done by the writer on the inorganic constituents of various plants.

[159]Scattered throughout the veterinary literature one finds cases of poisoning in animals with symptoms similar to those occurring in locoed animals which are attributed to eating plants grown on a peculiar soil, as in Oserow, Ueber Krankh. d. Pferde, welche Aehnlichkeit mit der Cerebro-spinal meningitis haben, aber durch Vergiftungen mit Gräsern von Salzgründen (Salzmooren) verursacht werden, Journ. f. Allgem. Veterinär-Medicin, St. Petersburg, p. 486, 1906. Abstract in Jahresber. über d. Leistungen auf dem Gebiete d. Veterinär-Medicin, vol. 26, p. 226, 1906.—Compare also Étude sur Quelques Plantes Vénéneuses des Regions Calcaires, Bul. Soc. Cent. de Méd. Vét., vol. 48, p. 378. 1894.

[160]After completing this work the writer found that Sayre had said that he "had the suggestion that the harm coming from this plant is due to the inorganic constituents; this clue has been followed up, but like the others has brought us no nearer to the solution of the problem." Kans. Acad. Sci. Trans., vol. 18, p. 144. 1903.

The filtrate from the ash from 200 grams of driedAstragalus mollissimus, from Imperial, Nebr., after similar treatment with acetic acid water and freed from free acid, killed a rabbit in several hours.

Hydrochloric acid also rendered the toxic agent of the ash soluble in water, but proved unsuitable for our work, as it was found impossible to obtain neutral residues by mere evaporation on the bath. At first one of the heavy metals or members of the H2S group[161]was suspected, but on passing H2S into the slightly acid extract of the ash noactive precipitate resulted, but the filtrate remained active.[162]A special Marsh test was, however, made for arsenic and antimony with negative results. A test for tungsten with zinc and hydrochloric acid proved negative.

Members of the ammonium sulphid group were then suspected, but while ammonium hydrate alone gave a heavy white precipitate, this precipitate, as also the black one with ammonium sulphid, proved inactive save when not thoroughly freed from acid (used for solution). The action of this ammonium sulphid precipitate on rabbits was watched for sixteen days, but without result. Nevertheless, the writer still suspected some of the rare earths.[163]

Sestini[164]had found that if certain plants were nourished with a solution of a beryllium salt, in the ash of these plants could be shown the presence of beryllium.

Two grams of beryllium chlorid were fed in aqueous solution to a rabbit weighing 1,800.2 grams. In four days this animal lost 241 grams and died. The stomach showed the same general pallor seen in chronic locoed rabbits, but no ulcers. The tests for beryllium by Sestini’s method, however, failed to show beryllium in the active loco plants examined.

Thorium chlorid, cerium chlorid, and lanthanum chlorid in 2-gram doses and zirconium chlorid in 3-gram doses produced no chronic symptoms in rabbits or, in fact, any disturbance. Titanium chlorid, 2.5 grams, evaporated in the air and then fed in an emulsion to a rabbit, also proved inactive, but this inactivity may have been due to its insolubility.

Thallium nitrate c. p., in aqueous solution, in 2-gram doses, killed a rabbit weighing 2,154.6 grams in two hours and fifteen minutes. The stomach in this case, while pink, was not hemorrhagic.

Zirconium chlorid has an astringent taste, and if fed repeatedly will cause the metallic astringent action. On boiling an acetic acid solution of the ash with sodium acetate a precipitate formed.[165]

The presence of zirconium was thus suspected and Dr. E. C. Sullivan, of the United States Geological Survey, estimated it to bepresent in the ash of a sample ofAragallus lambertiin about 0.01 per cent zirconium oxid, with also 0.1 per cent titanium dioxid.[166]

Zirconium chlorid, 3 grams, was fed in aqueous solution to a rabbit weighing 850.5 grams. This rabbit lost 96 grams in seven days, and was then fed 3 grams more of the same solution and the following day 2 grams more. It died eight days later, weighing 656 grams. The stomach and intestines were contracted, but showed no ulcers. However, 4 grams killed a rabbit in two hours and thirty-two minutes.

The filtrate, after treating an active solution of the ash with hydrogen peroxid, proved active, thus showing that zirconium was not entirely responsible for the poisonous action.

Yttrium, while not found in the plant, was administered as yttrium chlorid to a rabbit weighing 1,530 grams in 2-gram doses in solution. This animal gained 113.4 grams in five days.

Didymium chlorid c. p., in 3-gram doses, was fed to a rabbit weighing 1,020 grams. This rabbit lost 70 grams in four days.

The administration of manganese acetate[167]in 2-gram doses was followed by a gain in weight of a rabbit of 42.5 grams, while a dose of 3 grams killed a rabbit weighing 1,077 grams in two hours and thirty minutes. Wohlwill[168]has emphasized the fact that the members of the iron group owe their comparative harmlessness to not being absorbed by the gastro-intestinal tract.

No zinc was found in the plant.[169]

It is well recognized that potassium salts given hypodermically are decidedly toxic and that ammonium salts given per os will kill, so that the writer considered the possibility of other members of the group being responsible for the injurious action. The fact that the alkaline distillate of the plant proved inactive eliminated the ammonium salts.

Cæsium chlorid c. p., 2 grams, was fed in aqueous solution to a rabbit weighing 1,077.2 grams. In six days this animal lost 255 grams in weight, when it died.[170]

A second rabbit, weighing 1,020.5 grams, was fed with 2 grams of the same solution and lost 368 grams in twenty-one days. The spectroscopic test, however, failed to show cæsium in the ashed plant. Rubidium chlorid c. p., in 2-gram doses, proved inactive. The platinum chlorid precipitate from the extract of the plant proved inactive.

The fact that the filtrate after precipitation of the phosphates by tin and nitric acid and H2S was active excluded the phosphoric acid radical, and the filtrate after treatment with BaCO3and AgO being active excluded the H2SO4and HCl radicals as the toxic body. Fluorine was proved to be absent.

A radio-active substance was suspected, but Dr. L. J. Briggs, Physicist of Bureau of Plant Industry, reported that the dried plant showed no special amount of radio-activity.[171]

Power and Cambier, Sayre, and Kennedy had previously called attention to the abundance of calcium in the plant, and the writer’s investigations confirm this. Pharmacologists are averse to believing calcium given per os poisonous. The writer has, however, fed 5 grams of the acetate of calcium in solution to a rabbit weighing 652 grams. This animal died in two hours, with marked irritation of the stomach, the result being due to the so-called “salt action.” Much larger amounts were fed in divided doses, but without injury. Calcium phosphate and calcium sulphate in 2-gram doses proved harmless to a rabbit weighing about 1,400 grams. Three grams of magnesium acetate[172]were fed in solution for five successive days to a rabbit weighing 1,417 grams, but without apparent effect.

Strontium acetate c. p., in 2-gram doses, likewise caused no disturbance.[173]No strontium in any amount recognizable by chemical tests was proved in the plant. So that by a process of exclusion the writer was forced to think of barium as the main cause of the trouble.

The writer noted that if the ashed plant was extracted with H2SO4water and this extract freed from sulphuric acid with PbCO3and H2S the solution proved inactive to rabbits and also that after this extraction the acetic acid extract of the ash failed to kill. In other words, the sulphate of our body was insoluble in water. At times in passing H2S into active solutions of the ashed plant freed from the acetic acid by evaporation the filtrate and likewise the precipitate were inactive. Noyes and Bray[174]have noted that if H2S is passed into certain solutions in the presence of an oxydizing agent, such as ferric iron, H2SO4would be formed, which would throw any barium out of solution.

In one blood-pressure record made with a dog (vagi nerves cut), a rise in blood pressure (a characteristic physiological action of barium) was seen to follow the intravenous injection of the aqueous extract of the plant, in spite of its normal acid reaction.

Accidentally the writer found that Sprengel[175]had reported the presence of barium inAstragalus exscapus, a closely allied plant. Barium has also been found in the vegetable world by Scheele in 1788, and later by Eckard,[176]who found it in beech, while Forchhammer[177]proved it in birch, and Lutterkorth found it in the soil of the same area in which Eckard worked. Dworzak[178]noted the occurrence of traces of this element in wheat grown along the Nile, and Knop[179]found it in the soil. Doctor Balfour, of Khartum, Egypt, informed the writer that he knew of no cases in which this barium in wheat had produced poisoning. Hornberger[180]found barium both in the red beech grown in Germany and in the soil on which these trees grew. It has also been claimed that various marine plants may take up barium from the sea.[181]

Hillebrand[182]has called attention to the fact that the igneous rocks of the Rocky Mountains showed a higher percentage of barium than rock from other portions of the United States, so that under these conditions one might expect the presence of barium in plants growing in this region. A sample ofAragallus lambertiand one ofAstragalus mollissimuswere sent to the Bureau of Chemistry for spectroscopic examination for various elements and they reported traces of barium in each.[183]

With these arguments the writer felt sure of the presence of barium, and the matter was discussed with Dr. E. C. Sullivan, of the United States Geological Survey, and he kindly corroborated the conclusions reached as to the presence of barium, controlling its presence by means of the spectroscope, and estimated it roughly as 0.1 per cent BaO in the ash of a sample ofAragallus lamberti(6.3 milligrams BaSO4in 4 grams ash). This determination was made by Hillebrand’s method.

Kobert has anticipated this result, saying that “all plants are in the position occasionally to take up barium combinations from the soil,” and “the plants which thus contain barium may act injuriously to men and animals.”[184]

FOOTNOTES:[161]Swain, R. E., and Harkins, W. D. Arsenic in Vegetation Exposed to Smelter Smoke. Journ. Amer. Chem. Soc., vol. 30, p. 915. 1908.—Harkins, W. D., and Swain, R. E. The Chronic Arsenical Poisoning of Herbivorous Animals. Journ. Amer. Chem. Soc., vol. 30, p. 928, 1908.[162]A similar extract was sent to the Bureau of Chemistry, and that Bureau also reported an absence of the elements of the H2S group.[163]Bachem, C. Pharmakologisches über einige Edelerden. Arch. Internat. de Pharmacodyn., vol. 17, p. 363. 1907.[164]Sestini, F. Esper. di Vegetaz. del Frumento con Sostituz. della Glucina alla Magnesia. Staz. Sper. Agrar. Ital., vol. 20, p. 256. 1891.—Di alcuni Elementi Chimici Rari a Trovarsi nei Vegetabili. Staz. Sper. Agrar. Ital., vol. 15, p. 290. 1888.Note.—The ammonium sulphid precipitate was very small if the phosphates were first removed with tin and nitric acid.[165]Böhm, C. R. Darstellung d. seltenen Erden, vol. 1, p. 40. 1905.[166]Wait, C. E. Occurrence of Titanium. Journ. Amer. Chem. Soc., vol. 18, p. 402. 1896.Note.—There seem to be no records of any study of the pharmacological action of titanium.[167]Compare Jaksch, R. v. Ueber Mangantoxikosen und Manganophobie. Münch. Med. Woch., p. 969. 1907.[168]Wohlwill, F. Ueber d. Wirkung d. Metalle d. Nickelgruppe. Arch. f. Exper. Path., vol. 56, p. 409. 1907.[169]Laband, L. Zur Verbreitung des Zinkes im Pflanzenreiche. Zeits. f. Untersuch. d. Nahrungs u. Genussmittel, vol. 4, p. 489. 1901.[170]Cæsium occurs in various plants and the possibility of poisoning by this element must be considered. It is hoped that the writer may be able to undertake a more thorough pharmacological study of this element.[171]Acqua, C. Sull’accumulo di Sostanze Radioattive nei Vegetali. Atti della Reale Accad. dei Lincei, 5 s, vol. 16, sem. 2, p. 357. 1907.[172]Compare Meltzer, S. J. Toxicity of Magnesium Nitrate When Given by Mouth. Science, vol. 26, p. 473. 1907.[173]Burgassi, G. Modificaz. del Ricambio per Azione dello Stronzio. Archiv. di Farmacol., vol. 6, p. 551. 1907.[174]Noyes, A. A., and Bray, W. C. System of Qualitative Analysis for the Common Elements. Journ. Amer. Chem. Soc., vol. 29, pp. 168, 172, and 191. 1907.Note.—Barium sulphate is nontoxic on account of its insolubility. Orfila fed 16-24 grams to dogs without causing any disturbance. Bary, A. Beitr. z. Baryumwirkung. Dorpat, 1888, p. 25.[175]Sprengel, C. Von den Substanzen der Ackerbrume und des Untergrundes, Journ. f. Techn. u. Œkon. Chem., vol. 3, p. 313. 1828.[176]Eckard, G. E. Baryt, ein Bestandtheil der Asche des Buchenholzes. Annal. der Chem. u. Pharm., n. s., vol. 23, p. 294. 1856.[177]Forchhammer, J. G. Ueber den Einfluss des Kochsalzes auf die Bildung der Mineralien. Annal. d. Physik u. Chemie, vol. 5, p. 91. 1905.—Lutterkorth, H. Kohlensäurer Baryt, ein Bestandtheil des Sandsteines in der Gegend von Göttingen. Annal. d. Chem. u. Pharm., n. s., vol. 23, p. 296. 1856.[178]Dworzak, H. Baryt unter den Aschenbestandtheilen des. Ægyptischen Weizen. Landw. Versuchs.-Stat., vol. 17, p. 398. 1874.[179]Knop, W. Analysen von Nilabsatz. Landw. Versuchs.-Stat., vol. 17, p. 65. 1874.—Compare also Demoussy, E., Absorption par les Plantes de Quelques Sels Solubles, Thése, Paris, 1899.—Knop, W., Einige neue Resultate der Untersuchung über die Ernährung der Pflanze, Ber. ü. Verhandl. d. königl. sächs. Gesells. d. Wissens. zu Leipzig, Math. Phys. Cl., vol. 29, p. 113, 1877.—Suzuki, U., Can Strontium and Barium Replace Calcium in Phænogams? Bul. Coll. Agric. Tokio Imp. Univ., vol. 4, p. 69, 1900-1902.[180]Hornberger, R. Ueber d. Vorkommen d. Baryums in d. Pflanze und im Boden. Landw. Versuchs.-Stat., vol. 51, p. 473. 1899.[181]Roscoe, H. E., and Schorlemmer, C. Treatise on Chemistry, vol. 2, p. 455. 1897.[182]Hillebrand, W. F. Analysis of Silicate and Carbonate Rocks. Dept. Interior, U. S. Geol. Survey, Bul. 305, p. 18. 1907.[183]This report came from the Plant Analysis Laboratory of the Bureau of Chemistry, a different one from that which later controlled the writer’s tests quantitatively and qualitatively. In other words, the conclusions of the writer as to the presence of barium were controlled by three separate individuals.[184]Kobert, R. Kann ein in einem Pflanzenpulver gefundener abnorm höher Barytgehalt erklärt werden durch direkte Aufnahme von Baryumsalze durch die lebende Pflanze aus dem Boden? Chem. Zeit., vol. 10, p. 491. 1899.Note.—The writer has also found barium in entirely different botanical families from the loco-weed, and it is hoped a report can shortly be made of some of these.Note.—The first sample of ash analyzed by the Bureau of Chemistry had 0.21 per cent Fe2O3, 0.92 per cent Al2O3, 0.98 per cent CaO, 0.37 per cent MgO, 5.50 per cent SiO2. The second lot was only examined for certain constituents, and gave K2O, 2.25 per cent; CaO, 1.20 per cent; MgO, 0.41 per cent; P2O5, 0.52 per cent; and SO3, 0.24 per cent.

FOOTNOTES:

[161]Swain, R. E., and Harkins, W. D. Arsenic in Vegetation Exposed to Smelter Smoke. Journ. Amer. Chem. Soc., vol. 30, p. 915. 1908.—Harkins, W. D., and Swain, R. E. The Chronic Arsenical Poisoning of Herbivorous Animals. Journ. Amer. Chem. Soc., vol. 30, p. 928, 1908.

[162]A similar extract was sent to the Bureau of Chemistry, and that Bureau also reported an absence of the elements of the H2S group.

[163]Bachem, C. Pharmakologisches über einige Edelerden. Arch. Internat. de Pharmacodyn., vol. 17, p. 363. 1907.

[164]Sestini, F. Esper. di Vegetaz. del Frumento con Sostituz. della Glucina alla Magnesia. Staz. Sper. Agrar. Ital., vol. 20, p. 256. 1891.—Di alcuni Elementi Chimici Rari a Trovarsi nei Vegetabili. Staz. Sper. Agrar. Ital., vol. 15, p. 290. 1888.Note.—The ammonium sulphid precipitate was very small if the phosphates were first removed with tin and nitric acid.

Note.—The ammonium sulphid precipitate was very small if the phosphates were first removed with tin and nitric acid.

[165]Böhm, C. R. Darstellung d. seltenen Erden, vol. 1, p. 40. 1905.

[166]Wait, C. E. Occurrence of Titanium. Journ. Amer. Chem. Soc., vol. 18, p. 402. 1896.Note.—There seem to be no records of any study of the pharmacological action of titanium.

[166]Wait, C. E. Occurrence of Titanium. Journ. Amer. Chem. Soc., vol. 18, p. 402. 1896.

Note.—There seem to be no records of any study of the pharmacological action of titanium.

[167]Compare Jaksch, R. v. Ueber Mangantoxikosen und Manganophobie. Münch. Med. Woch., p. 969. 1907.

[168]Wohlwill, F. Ueber d. Wirkung d. Metalle d. Nickelgruppe. Arch. f. Exper. Path., vol. 56, p. 409. 1907.

[169]Laband, L. Zur Verbreitung des Zinkes im Pflanzenreiche. Zeits. f. Untersuch. d. Nahrungs u. Genussmittel, vol. 4, p. 489. 1901.

[170]Cæsium occurs in various plants and the possibility of poisoning by this element must be considered. It is hoped that the writer may be able to undertake a more thorough pharmacological study of this element.

[171]Acqua, C. Sull’accumulo di Sostanze Radioattive nei Vegetali. Atti della Reale Accad. dei Lincei, 5 s, vol. 16, sem. 2, p. 357. 1907.

[172]Compare Meltzer, S. J. Toxicity of Magnesium Nitrate When Given by Mouth. Science, vol. 26, p. 473. 1907.

[173]Burgassi, G. Modificaz. del Ricambio per Azione dello Stronzio. Archiv. di Farmacol., vol. 6, p. 551. 1907.

[174]Noyes, A. A., and Bray, W. C. System of Qualitative Analysis for the Common Elements. Journ. Amer. Chem. Soc., vol. 29, pp. 168, 172, and 191. 1907.Note.—Barium sulphate is nontoxic on account of its insolubility. Orfila fed 16-24 grams to dogs without causing any disturbance. Bary, A. Beitr. z. Baryumwirkung. Dorpat, 1888, p. 25.

[174]Noyes, A. A., and Bray, W. C. System of Qualitative Analysis for the Common Elements. Journ. Amer. Chem. Soc., vol. 29, pp. 168, 172, and 191. 1907.

Note.—Barium sulphate is nontoxic on account of its insolubility. Orfila fed 16-24 grams to dogs without causing any disturbance. Bary, A. Beitr. z. Baryumwirkung. Dorpat, 1888, p. 25.

[175]Sprengel, C. Von den Substanzen der Ackerbrume und des Untergrundes, Journ. f. Techn. u. Œkon. Chem., vol. 3, p. 313. 1828.

[176]Eckard, G. E. Baryt, ein Bestandtheil der Asche des Buchenholzes. Annal. der Chem. u. Pharm., n. s., vol. 23, p. 294. 1856.

[177]Forchhammer, J. G. Ueber den Einfluss des Kochsalzes auf die Bildung der Mineralien. Annal. d. Physik u. Chemie, vol. 5, p. 91. 1905.—Lutterkorth, H. Kohlensäurer Baryt, ein Bestandtheil des Sandsteines in der Gegend von Göttingen. Annal. d. Chem. u. Pharm., n. s., vol. 23, p. 296. 1856.

[178]Dworzak, H. Baryt unter den Aschenbestandtheilen des. Ægyptischen Weizen. Landw. Versuchs.-Stat., vol. 17, p. 398. 1874.

[179]Knop, W. Analysen von Nilabsatz. Landw. Versuchs.-Stat., vol. 17, p. 65. 1874.—Compare also Demoussy, E., Absorption par les Plantes de Quelques Sels Solubles, Thése, Paris, 1899.—Knop, W., Einige neue Resultate der Untersuchung über die Ernährung der Pflanze, Ber. ü. Verhandl. d. königl. sächs. Gesells. d. Wissens. zu Leipzig, Math. Phys. Cl., vol. 29, p. 113, 1877.—Suzuki, U., Can Strontium and Barium Replace Calcium in Phænogams? Bul. Coll. Agric. Tokio Imp. Univ., vol. 4, p. 69, 1900-1902.

[180]Hornberger, R. Ueber d. Vorkommen d. Baryums in d. Pflanze und im Boden. Landw. Versuchs.-Stat., vol. 51, p. 473. 1899.

[181]Roscoe, H. E., and Schorlemmer, C. Treatise on Chemistry, vol. 2, p. 455. 1897.

[182]Hillebrand, W. F. Analysis of Silicate and Carbonate Rocks. Dept. Interior, U. S. Geol. Survey, Bul. 305, p. 18. 1907.

[183]This report came from the Plant Analysis Laboratory of the Bureau of Chemistry, a different one from that which later controlled the writer’s tests quantitatively and qualitatively. In other words, the conclusions of the writer as to the presence of barium were controlled by three separate individuals.

[184]Kobert, R. Kann ein in einem Pflanzenpulver gefundener abnorm höher Barytgehalt erklärt werden durch direkte Aufnahme von Baryumsalze durch die lebende Pflanze aus dem Boden? Chem. Zeit., vol. 10, p. 491. 1899.Note.—The writer has also found barium in entirely different botanical families from the loco-weed, and it is hoped a report can shortly be made of some of these.Note.—The first sample of ash analyzed by the Bureau of Chemistry had 0.21 per cent Fe2O3, 0.92 per cent Al2O3, 0.98 per cent CaO, 0.37 per cent MgO, 5.50 per cent SiO2. The second lot was only examined for certain constituents, and gave K2O, 2.25 per cent; CaO, 1.20 per cent; MgO, 0.41 per cent; P2O5, 0.52 per cent; and SO3, 0.24 per cent.

Note.—The writer has also found barium in entirely different botanical families from the loco-weed, and it is hoped a report can shortly be made of some of these.

Note.—The first sample of ash analyzed by the Bureau of Chemistry had 0.21 per cent Fe2O3, 0.92 per cent Al2O3, 0.98 per cent CaO, 0.37 per cent MgO, 5.50 per cent SiO2. The second lot was only examined for certain constituents, and gave K2O, 2.25 per cent; CaO, 1.20 per cent; MgO, 0.41 per cent; P2O5, 0.52 per cent; and SO3, 0.24 per cent.

The reports of the ash analyses of the loco plants show marked variations in the total amount of the ash. Thus, fromAragallus lambertiDyrenforth obtained 4.32 per cent and O’Brine 13.52 per cent of ash. The Bureau of Chemistry analyzed two different samples of this dried plant and reported in one case 11.15 per cent and in the second 11.64 per cent of ash. O’Brine[185]obtained 13.52per cent of ash from the same species. The writer’s analysis[186]gave in one sample ofAragallus lamberti, collected at Hugo, Colo., in 1907, 18.8 per cent of ash; a second lot (1907), 12.44 per cent; a third (1906), 11 per cent, and a fourth (May, 1905) gave 37.3 per cent of ash.[187]One lot from Woodland Park, Colo. (October, 1906), gave 6.4 per cent. One lot from Hugo, Colo. (October, 1907), yielded 9.6 per cent.

In the case ofAstragalus mollissimus, Wentz obtained 6.76 per cent, Sayre 12.01 per cent, Kennedy 20 per cent, O’Brine 12.15 per cent, while the sample analyzed by the Bureau of Chemistry gave 18.4 per cent of ash. One sample from Kit Carson County, Colo. (December, 1906), which proved inactive physiologically, gave an ash content of 6.9 per cent. A sample ofAstragalus missouriensiscollected at Hugo, Colo., June, 1907, yielded an ash content of 21.8 per cent, and anAstragalus missouriensiscollected at Pierre, S. Dak., September, 1907, yielded 27 per cent. AnAstragalus nitidusfrom Custer, S. Dak. (July, 1907), gave 5.2 per cent ash, while anAstragalus nitiduscollected at Woodland Park, Colo., in October, 1906, yielded 7.8 per cent, and another specimen ofAstragalus nitidusalso collected at Woodland Park, Colo., in October, 1907, gave 12.2 per cent. AnAstragalus drummondiifrom Custer, S. Dak. (July, 1907), gave 5.9 per cent.Astragalus pectinatus(Hugo, June, 1907) yielded 6.1 per cent. A fresh (undried) specimen ofAstragalus mollissimus(unknown origin, November, 1907) yielded 3.8 per cent of ash. One sample ofAstragalus decumbens(Ephraim, Utah, August, 1907) gave 21.8 per cent of ash.

These determinations must necessarily be only approximate, as the plants were collected by different persons who exercised different degrees of care in freeing them from adherent soil, and possibly in drying the plants, so that the main value of these figures is their aid in determining the amount of barium present.

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