Art. XXXIII.—A. Contribution to our Knowledge of the Physiological Action of Tutin.*
[Read before the Otago Institute, 10th November, 1908.]
|1. General and descriptive||287|
|2. Botany of the tutu plant||291|
|3. Work of previous observers||293|
|4. The active principle||294|
|B. Original Work.|
|1. General action of tutin on mammals—|
|2. Action on birds||299|
|3. Action on reptiles||304|
|4. Action on amphibia||304|
|5. Action on fishes—|
|(a.) General action||306|
|(b.) Comparison of action of tutin and pierotoxin||309|
|(c.) Effects of tutin hydrolysed by hydrochloric acid||311|
|(d.) Action of alkalies on tutin||313|
|(e.) Influence of “surviving” organs||314|
|6. Action on insects||314|
|7. Action on larvae||315|
|8. Action on molluscs||315|
|9. Action on infusoria, and amcebae||315|
|10. Action on putrefactive bacteria||316|
|11. Action on yeast||316|
|12. Action on germination of seeds||317|
|13. Action on tissues—|
|(a.) Action on primitive protoplasm||317|
|(b.) Ciliated epithelium||317|
|(c.) Action on striped muscle and nerve-terminations||318|
|14. Action on the different systems—|
|(a.) Alimentary system||319|
|(b.) Etemopoietic system||320|
|(c.) Circulatory and respiratory systems||320|
|(d.) Urinary system||323|
|(e.) Genital system||323|
|(f.) Nervous system||323|
|(g.) General nutrition||326|
|15. Fate of tutin in the body||327|
|16. Immunity or tolerance||327|
|17. Pharmacological relationship of tutin||328|
|18. Action of remedies||329|
|19. General summary||332|
|Protocols of experiments||333|
[Footnote] * Being a thesis presented to the University of Edinburgh, for which the degree of Doctor of Medicine was granted.
1. General and Descriptive.
On the 20th May, 1773, at Queen Charlotte Sound, Captain. Cook sent on shore a ewe and a ram which he had brought from the Cape of Good Hope with a view to stocking New Zealand with sheep. On the 22nd he “received the unpleasant intelligence that the ewe and ram which with so much care and trouble he had brought to this place were both of them, found dead. It was supposed that they had eaten some poisonous plant, and by this accident all the Captain's hopes of stocking New Zealand with a breed of sheep were instantly blasted” (1). He tried again, however, with goats, but with little better result.
Cook's supposition that the flora of New Zealand includes a plant possessing highly toxic properties was well founded. That his ewe and ram shared the fate that has since been meted out to many thousands of sheep and cattle, and to not a few human beings, is in the highest degree probable.
The poisonous plant whose existence was suspected in 1773 has long since been identified as a Coriaria, and is known throughout the Dominion by its Maori name tutu, or, as Europeans often pronounce it, “toot.”
The difficulties met with by Cook in his attempts to stock the country were again encountered by the early settlers. Large numbers of their flocks and herds were destroyed by eating the leaves and succulent young shoots of this attractive shrub, for it abounded everywhere, and grew most profusely where the soil, by its richness, offered an inducement to the pioneer to settle.
Animals hungry and in poor condition were particularly prone to succumb to the effects of the poison; and, as these conditions prevailed with most of the beasts landed from the ships, it will be understood how great an impediment to stocking the country this noxious plant proved. The very first issue of the Lyttelton Times (2) notices the death of three out of five cows that had just been landed—one fell over a cliff, and two others were poisoned by eating tutu. The newspaper, in warning settlers of the dangerous properties of the plant, says, “It is impossible to take too much care in landing cattle at this place. To beasts just out of a ship the tutu, of which there is abundance here, is certainly fatal.”
The damage done to stock was enormous, as may be gathered from the following quotation from Dr. Lauder Lindsay's article “On the Toot Plant and Poison of New Zealand” (3): “In the course of a tour through the New Zealand provinces during the latter part of 1861 and earlier months of 1862 I was everywhere struck by the abundant evidences of devastation produced among flocks and herds from their feeding on the ‘toot’ plant, one of the most widely distributed and familiar indigenous shrubs of the country. One settler friend told me of his having lost by ‘tooting’ two hundred and fifty sheep; another, eighty to a hundred sheep of a flock of four hundred; a third, seven of sixteen bullocks; a fourth, six of twenty-four cattle; a fifth, twenty-four cattle; a sixth, six of eight cattle—each of these instances in a single night. Another flock-master lost four hundred sheep out of a flock of two thousand, twenty-five being frequently dead of a night. In other words, he seemed a fortunate farmer or runholder who had not lost more than 25 per cent., or one-fourth, of his stock from toot poisoning; while in some instances the losses were so high as 75 per cent., or three-fourths. Some of the colonists had suffered so severely from losses of bullocks by toot poisoning that they were at the trouble and expense
of attaching a boy to each of their bullock-teams, solely for the purpose of preventing their animals-feeding on this pest of the colony. Such incidents I found were of daily occurrence. I met few settlers who had not at some period had occasion from this cause to mourn the loss of sheep or bullocks—the former sometimes by the hundred, the latter by the dozen.” These remarks of Lindsay serve well to illustrate the deadly nature of the plant, and the embarrassment it offered to the pioneers of the country. They also point to the importance and great desirability of an investigation of the physiological action of the poison being made, in order that the treatment of its effects may be conducted on rational lines, and as the first step towards the discovery of an appropriate antidote.
But while sheep and cattle have been the chief victims, human beings have not proved exempt, and he was a lucky farmer who lost only his cows or his sheep. Too often one of his children succumbed to the effects of eating the berries or young shoots. Nor is this strange, for the shrub in full fruit is a very striking and attractive object. The numerous racemes of richly coloured, tempting-looking berries — at a glance not unlike black currants—could hardly be overlooked or neglected by the child or thirsty traveller ignorant of their dangerous properties.
Numerous cases of poisoning must have occurred, but the recorded deaths from this cause are not many. The following are all that occur in the literature that has been laid under contribution: 1. Thomson (4), in 1859, mentions that up till that date several children had died from eating the berries. 2. Lauder Lindsay (3) mentions the case of twelve French sailors who were poisoned by eating the berry; four of them died. 3. The Otago Colonist (5) records the death of one of two children in 1861 who had eaten the shoots. 4. The Otago Daily Times notices the death of a young man in 1862 from eating the shoots (6). 5. Easterfield and Aston (7) put on record the following cases: A girl in 1854–55, from eating the berries; a boy in 1860, from eating the berries; two cases from eating the berries—one died, the other recovered, with impaired memory.
For the purposes of this paper the writer asked of the Registrar-General a return of all cases of death from tutu poisoning that occur in the records. In his reply the Registrar-General stated that he was unable to make a return, as cases of this kind are classed in the vital statistics under the general heading “Accidental Poisoning.” He had had the statistical tables relating to inquests examined for thirty years back, and found that only four deaths occurred from eating poisonous berries, one in each of the years 1889, 1891, 1896, and 1902. It is probable that the berry in each of these cases was the tutu-berry, but in only one case—that of 1889—was it stated to be so.
Effect of the Plant on Animals.
Horses.—Statements vary as to the effect on horses. It is said that they have been known to eat freely of the plant without injury (8); and, again, that C. thymifolia is highly poisonous to them (9). They are said to refuse the young shoots, but have been known to eat the berries (3). If they escape, it is probably because they do not eat enough of the plant; that they eat freely without evil result is not credible.
Birds are regarded as immune. They certainly eat freely of the berries without, ill effect. The question of their immunity will be considered later.
Rabbits are said to be immune (10), (11), and certainly the tutu does not seem to have checked their increase. They probably do not eat the plant.
Elephants.—Sir Julius von Haast (12) records the death of an elephant from tutu-poisoning. The animal was marched inlands by its owner for a considerable distance, and on arriving at a suitable halting-place, where the vegetation was abundant, was allowed to feed. The grass had been burnt off during the previous season, and had shot up again, together with a large crop of tutu-shoots. The elephant fed for four hours, and then drank freely from a neighbouring stream. It then began to reel, fell on the ground, and died in three hours.
Sheep and Cattle.—The following extract from a letter received from the manager of a large sheep-station gives an excellent account of the effects of tutu upon sheep: “The effect on sheep is that they will stand still, trembling as if palsied, froth at the mouth, with their jaws going continually, and their teeth grinding. Suddenly they will fall over, with their limbs rigid, as if suffering from strychnine poisoning. If assisted on, to their legs they are absolutely mad, and will rush against a fence or over a precipice, and will pay no attention to man, or dog, or animals of their own kind. With animals that have eaten less of the plant, symptoms do not appear unless they are disturbed, and then the effect is shown with terrible suddenness; a bark from a dog or a sudden run for a few yards will be almost certain to start the poison to work. In cases like the above, however, the affected sheep' generally recover if left alone.” In their wild career they often injure themselves against obstacles, or rush into creeks and are drowned. More frequently they die in convulsions.
Cattle are similarly affected, but the wild delirium is even more marked, in their case. Popularly they are said to go mad; and the wild way in which they wheel round and round, gallop aimlessly about, kicking, charging, and rushing blindly against rocks and other obstacles, lends colour to the popular opinion. The wild career continues until the animal, overcome by exhaustion, falls to the ground, becomes comatose, and dies in convulsions.
In the light of this account, the symptoms displayed by one of Captain Cook's animals is interesting: “The ram was taken with fits, bordering, on madness… One night he was seized with one of these, fits and ran headlong into the sea, but soon came out again, and seemed quite easy. Presently after he was seized with another fit, and ran along the beach … and was never seen more” (13).
Animals are frequently found distended with gas, “blown” after death. This is probably due to rapid fermentation of the leaves ingested, and is similar to the condition met with in cattle after eating freely of clover. As with clover, it is more pronounced when the tutu is eaten wet.
Effect on Human Beings.
The symptoms of poisoning by the plant in the human subject include vomiting, giddiness, delirium, great excitement, stupor, coma, and convulsions.
In a fatal case reported in the Otago Colonist of the 25th October, 1861, the physician who attended the case stated in his evidence at the inquest that he found the child perfectly pale, with teeth clenched. The breathing was difficult, the lips livid, and the pupils much dilated. For about five minutes the rigidity went off, and the pupils contracted; but a relapse occurred, the teeth were set again, and the child gradually sank back, “without any symptom of convulsion or suffering.” In this case it was the
young shoots that had been eaten. The jury appended the following rider to their verdict: “The jury would recommend the Provincial Government to keep up a standing advertisement in the public prints warning persons of the poisonous nature of the tutu plant, the young sprouts as well as the berries, and also giving descriptions of the same” (5).
Lauder Lindsay (3) records several cases that had been reported to him. In one case about half a pint of berries were eaten shortly after the evening meal. No effect was produced until 6 o'clock next morning, when, on attempting to rise as usual, the subject suddenly lost consciousness till 11 a.m. He was then conscious for a minute or two, but almost immediately relapsed into stupor, which continued for about twenty-four hours. When he came to himself he had lost his memory, and for half a day was unable to say where he was, or what he had been doing for the previous forty-eight hours. He gradually recovered, and there were no subsequent bad effects. Throughout the illness he had had no pain.
In another case two young men partook of some tutu-berries, about 4 p.m. One swallowed the berries, the other sucked them but spat out the seeds. The latter was unaffected, but the former was seized with convulsions about 9 p.m. The convulsions continued for about forty minutes, and then the patient gradually passed into a state of coma, which continued all next day. On recovering consciousness he was utterly oblivious of the particulars of his illness, and denied having eaten the berries. He was drowsy for a few days, but gradually recovered.
In a third case, retching, vomiting, and convulsions were the prominent symptoms. This case did not recover completely. A mental change, which was permanent, remained after the acute symptoms had subsided.
In a fourth case, in which particulars were given by Dr. Stewart, of Tuapeka Hospital, insomnia was one of the earliest symptoms. This was followed by tonic and clonic spasms, with coma, lasting for two days. During recovery, entire loss of memory was observed. The patients—there were two—did not know where they were, why they had been brought to hospital, or what their occupation was. They did not recollect having eaten tutu.
In a case noted by Dr. Hocken, the symptoms included tonic and clonic spasms lasting two days, coma, and loss of memory.
Considering the highly toxic nature of the plant, it is surprising that so few deaths from poisoning are on record. In the early days probably many fatal cases occurred of which no record was made. Nowadays cases of poisoning are rare. This may possibly be accounted for by the very distinctive appearance of the plant and the widespread knowledge of its poisonous nature. No one who has once seen tutu is likely to mistake it for anything else, and every school-child knows that “toot” is poisonous. Moreover, near the large centres of population the plant is not often now met with in places readily accessible to young children.
Among flocks and herds the mortality has also fallen, not because, as is sometimes supposed, the animals have acquired immunity or tolerance, but because in the more highly cultivated districts tutu has been largely exterminated. Sheep and cattle when moved from place to place are now sent by rail rather than driven. This alone has considerably reduced the mortality, for driven animals are particularly prone to suffer, and tutu is the commonest roadside plant in many districts.
The farmer, too, taught by experience, has become learned in the ways of managing his stock in relation to tutu. He takes care to avoid “toot”
country in the early spring, when the too tempting and especially toxic young sprouts are appearing. If sheep are to be turned out where tutu abounds they are first fed freely on English grass, for a well-fed sheep only nibbles tutu as he happens to meet it, and escapes; while a hungry sheep, finding it difficult to resist so ready a means of appeasing its appetite, fills its stomach, and succumbs. Indeed, it is a constant observation of farmers that tutu introduced into an empty stomach is more lethal than when taken into a stomach that already contains food. Cross-breds are said to be less readily affected than the more active and restless merino, and are therefore selected for distribution to tutu-infested areas.
But, in spite of these precautions, accidents, through, the ignorance of a shepherd, or the chance of an open gate, occasionally happen, and a heavy loss is experienced, as witness the following instance reported in 1905: “A settler left sixty bullocks about four years old in a field of swedes. In his absence they escaped from the paddock into a gully full of tutu (C. ruscifolia), where, on his return, he found forty-three of them dead. Mr. Clayton found the rumen packed with tutu leaves and branchlets” (14).
The tutu plant belongs to the natural order Coriariece, a small order of very doubtful relationship possessing but a single genus, Coriaria. The genus includes some eight or ten species, and has a rather remarkable distribution, species being met with in south Europe, South America, China, Japan, north Africa, India (Himalayas), and New Zealand.
The European species, C. myrtifolia, is well known to possess toxic properties. Its leaves have been used to adulterate senna with fatal effect, and numbers of cases of death are recorded from eating the berries. In 1862 several persons were said to be poisoned by eating snails that had been fattened on its leaves and young shoots (15). The symptoms of poisoning include vomiting and convulsions, and, on the whole, closely resemble those of tutu poisoning. In 1863 Riban (15) investigated the chemistry of this species, and separated a glucoside which he named “coriamyrtin.” The physiological properties of this compound will be referred to later.
The Himalayan species, C. nepalensis, is stated to be non-toxic, but, as the same has been said both of tutu and of C. myrtifolia, the statement must be accepted with reserve. The fruit is said to be eaten with impunity.
The American species, C. thymifolia, and the New Zealand species are said to be identical, and this statement has been used to prove a former land-connection between the two countries. It is more likely, however, that the order is a very ancient one, which has died out everywhere except in those places in which it is now found (8). Moreover, the identity of the two has been questioned.
The species met with in New Zealand are given by Cheeseman (16) as three in number—(1) C. ruscifolia, (2) C. thymifolia, (3) C. angustissima. The first is known locally as the “tree-toot,” the second as the “ground-toot,” and to both the name “tutu” applies. The Maoris have no name for C. angustissima.
There seems to be some division of opinion as to whether these three really constitute separate naturally distinct species, or whether the two last are merely varieties of the first. Lauder Lindsay, who in 1868 described, though with hesitation, four species—(1) C. arborea, (2) C. tutu, (3) C. thymifolia, (4) C. angustissima—says, “If only typical species be examined the
student will have little difficulty in accepting the foregoing as good species well distinguished from each other by habit, but if he extend his observations to … forms in the living state over wide areas, he will not fail to find them connected by transition states which he will frequently be puzzled to refer to one book species rather than another, partaking as they do of the characters of two or more of these species” (17); and he suggests that it might be preferable to regard them as mere forms of a most variable single type.
Cheeseman says of C. thymifolia that in its ordinary state it is distinct enough, but that the large-leaved forms pass directly into C. ruscifolia, and narrow-leaved varieties into C. angustissima (16).
G. M. Thomson (18) regards C. angustissima as a mere altitudinal variety of C. thymifolia.
The botanical characters of C. ruscifolia, as given by Cheeseman, are: “A shrub or small tree with spreading 4-angled branches, very variable in height and degree of robustness, sometimes attaining 25 ft. with a trunk 10 in. diameter, at others not more than 2–4ft., with almost herbaceous stems. Leaves 1–3 in., ovate or oblong-ovate, acute or acuminate, rounded or cordate at the base, sessile or very shortly petioled, 3–5-nerved. Racemes drooping, many-flowered, 4–12 in. long or more, slightly pubescent. Pedicels slender, ½–½ in., bracteolate at the base. Flowers small, green, ⅛–⅙ in. diameter, strongly proterogynous. Sepals broadly ovate, subacute. Filaments elongating after fertilisation. Fruit globose, purplish-black, of 5–8 cocci, enveloped by the persistent enlarged juicy petals” (16).
The herbaceous-stemmed shrub form of this is the more common. It takes this form in the open country and where the soil is dry. The roots creep and interlace below the surface, and in the spring stems shoot up from every part of the root, sometimes forming an almost impenetrable jungle. The stems may grow 10 ft. to 15 ft. in a single season. The tree form often grows solitary in the bush that lines the banks of streams. Shelter and moisture seem necessary to the attainment of this form.
C. ruscifolia occurs abundantly in all three islands of New Zealand, the Kermadee Islands, and the Chatham Islands, and is met with from sea-level to a height of 3,500 ft. C thymifolia occurs only in the North and South Islands of New Zealand; and C. angustissima is still more restricted in its distribution, occurring only in subalpine localities in the Provinces of Otago and Canterbury. These two species differ from the former chiefly in the size of the plant and of the leaves, and in their annual habit. All three forms are met with in abundance in the immediate neighbourhood of Dunedin. The tree form of C. ruscifoha may be found in the bush that lines the banks of the Water of Leith and its tributaries; while C. thymifolia and C. angustissima are plentiful on the hills that encircle the town.
All parts of the plant are poisonous, but the young shoots are more toxic than the leaves and fruit. The same has been noted of C. myrtifolia (15).
In the case of human beings it is usually the so-called berry that is eaten, though the shoots too are sometimes eaten by children. Of the berry, only the seed is poisonous. The strained juice is harmless, and from it the Maoris and early settlers made a non-intoxicating wine that was drunk in large quantities (19). Indeed, in the very early days the tutu was known as the wine-berry shrub. This wine, however, has not always proved to be above suspicion. Canon Stack (20) relates that on one occasion after partaking of some tutu-wine he was seized with alarming symptoms; he lost all feeling
in his extremities, a mist came over everything, and he thought that he was poisoned. The symptoms soon passed off, however, and he was none the worse.
Cattle and sheep are especially fond of the young, tender, asparagus-like shoots, but they also eat the leaves and branchlets with readiness.
3. Work of Previous Observers.
When one considers the harmful influence that this noxious plant has had upon the development of the country, it is remarkable that until recent years little effort was made to determine the nature of the poison.
In 1869 Skey (21) investigated the chemistry of C. ruscifolia. He showed that the poisonous principle is not an alkaloid, as was commonly thought, and with ether extracted from the seeds a green-coloured oil, 5 minims of which when given to a cat quickly produced the symptoms characteristic of tutu poisoning. Its highly toxic nature, together with certain peculiar chemical properties possessed by it, inclined him to the opinion that this oil was the active principle.
A year later, 1870, Hughes (10) attempted to separate an alkaloid, using the ground-shoots, and did indeed succeed in obtaining a crystalline substance, but failed to identify it. He thought that a heavy olive-coloured oily fluid which he also obtained and proved to be toxic might be the active principle and “a liquid alkaloid similar to conia.” He showed that lime destroyed the activity of the poison, and advocated its use as an antidote. In conjunction with Dr. Acheson, he conducted a series of experiments on cats and dogs, but more with the object of proving the toxicity of his extracts, and of determining the value of lime as an antidote, than with any idea of advancing our knowledge of the physiological action of the poison.
Hughes's results were adversely criticized by Skey (22), who held that the temperature used in Hughes's experiments was such as must have produced many side-products by destructive distillation, and among others acetate of ammonia, the presence of which, he thought, would sufficiently account for the reactions attributed by Hughes to the presence of an alkaloid.
Twenty years elapsed before any further investigation was undertaken, and then, in 1890, W. L. Christie (11) examined the physiological action of the oil that had first been extracted by Skeey. He made a series of experiments on mammals, including one upon himself, and briefly summarised the conclusions he arrived at as follows: (1) That tutu acts on the nerve-centres after absorption into the blood; (2) that the grey matter of the motor cortex is the part chiefly affected, and that this peripheral action (sic) causes epileptiform convulsions; (3) that vomiting is chiefly due to central causes, and that by its means, and perhaps by the renal secretion, the poison is removed from the body; and, lastly, (4) that dyspncea is due to poisoning of the respiratory centres, and when death ensues it is due to asphyxia from this cause or tetanus of the respiratory muscles—both may however, I believe, occur from coma.”
The chief interest in Christie's work, however, lies in the experiment upon himself. He took, in all, 9 grains of an extract made from the leaves gathered in the spring. He calculates that each grain of extract represents 100 grains of leaf; but, in the absence of data regarding the amount of tutin in tutu-leaves at different times of the year, and of details as to the exact
method of making the extract, it is impossible to calculate what dose of tutin was taken in this case. The first dose (4 ½ grains) was taken at 2.20 on Friday afternoon, and a second dose of the same amount at 4 p.m. An hour or two later he felt sick and faint, and began to vomit. The vomiting occurred at frequent intervals, and continued for twenty-four hours. At 8 p.m. he felt slight twitches an the legs and arms; and at 10.40 p.m. the medical student who was acting as clerk of the case noted that “all the muscles seemed to get tight, and there was foaming at the mouth.” At 10.50 the pulse-rate was 102 and the breathing heavy. Twenty minutes later the pulse was still 102, but the breathing was normal, and there was profuse perspiration. At 11.24 p.m. the clerk noted that “the subject spoke in a collected manner; getting right, but drowsy.” The vomiting continued till 8 p.m. on Saturday. The following day (Sunday) he felt sick and dull, but, though shaky, managed to attend to his duties. He states that sensation, was dulled and spirits below par for seven or eight days. For a month he was not in good tone, and then for the first time he felt a sensation of “pins and needles” in his fingers and toes, and felt the floor of his bedroom woolly when he rose in the morning. He could feel accurately with his fingers, but experienced a heavy, stiff, numb sensation when, he used them, and this symptom lasted a month.
4. The Active Principle.
The first substantial advance in the investigation of the chemical properties of tutu was made in 1900, when Easterfield and Aston (23) succeeded in isolating a crystalline glucoside, to which they gave the name “tutin.” All three species of Goriaria, were experimented upon, and crystals of tutin were obtained from each. The young shoots were found to yield a greater quantity (0.03 per cent.) than either the berries or the leaves:
Preparation: In the case of C. ruscifolia, “the fresh young shoots were finely divided, the juice expressed, filtered, and evaporated to a syrup nearly neutralised by carbonate of soda and shaken up with ether. The ether on evaporation deposited crystals of tutin. These were recrystallized from alcohol until the melting-point was constant.”
Properties: Tutin is described as a colourless, odourless, intensely bitter compound, which separates from alcohol in oblique-ended prisms, and from hot concentrated solutions in water in characteristic acicular forms. It is perceptibly volatile, sublimes readily at 120° to 130° C., and melts at 208° to 209° C. (uncorrected). It contains no nitrogen, and reduces Fehling solution after inversion by acid. The compound is therefore to be considered a glucoside, but the hydrolyzed substance yields with phenylhydrazine an amorphous precipitate which is not phenylglucosazone. Examination by Zeissl's method for methoxyl groups gave negative results. Strong sulphuric acid added to a few drops of a saturated aqueous solution of tutin gives a blood-red coloration.
Solubility: 100 grams water at 10° C. dissolve 1.9 grams tutin; 100 grams either at 10° C. dissolve 1.5 grams tutin; 100 grams alcohol at 10° C. dissolve 8.2 grams tutin. It is very soluble in acetone, sparingly soluble in chloroform, and soluble in benzine and carbon-disulphide.
The optical activity has been determined by Marshall. The substance is dextrorotatory, and the specific rotatory power is + 9.25. Easterfield and Aston found that when solutions of tutin were evaporated to dryness
with slaked lime the tutin underwent decomposition, and could not be recovered. This fact recalls Hughes's statement that lime destroyed the activity of the tutu poison.
The close relationship of the New Zealand tutu to the European C. myrtifolia, together with the fact that Lauder Lindsay (17) and others have thought it probable that the active principle of tutu is coriamyrtin, lends interest to the following table of differences existing between the two bodies, as given by Easterfield and Aston (23):—
|Melting-point||220° C. (according to Merck, 229° C.)||208° C.|
|Carbon||64.1 per cent.||60.7 per cent.|
|Hydrogen||6.6 per cent.||5.8 per cent.|
|With hydriodic acid and potash||Gives fuchsia-red colour||Gives nil.|
|Solubility in 100 parts of water||1.44 at 22° C.||1.9 at 10° C.|
|Effect on pupil||Contracts it||Dilates it.|
It may be said here, and will be shown later, that tutin has little, if any, action on the pupil when applied locally.
The physiological activity of the new compound was put to the test by Mr. Gilruth (23), then Chief Government Veterinarian in New Zealand. He administered to a pig weighing 17 kilograms, 0.129 grams of tutin, dissolved in water and mixed with half a pound of pollard. In half an hour uneasiness and spasmodic movements of the jaws were observed; then the breathing was noticed to be accelerated, and vomiting occurred. The symptoms gradually increased in severity, until the animal was seized with tetanic convulsions. The convulsions were at first separated by intervals of about fifteen minutes, but they gradually increased in severity and frequency until they became almost continuous. Each convulsive seizure, after beginning with a tonic spasm that lasted half a minute, was accompanied by screaming and stertorous breathing. Finally the animal died in a convulsion five hours after the poison had been administered.
Further experiments on cats testified to the marked toxicity of the compound—e.g., a dose of 0.01 gram killed a kitten weighing 1 kilogram in forty minutes; 0.001 gram administered to a cat weighing 2 kilograms caused a fit in three hours and illness for the next twenty-four hours. The same cat was afterwards killed by a dose of 0.003 gram. A dose of about a milligram caused nausea, vomiting, and incapacity for work for twenty-four hours in a full-grown, healthy man.
B. Original Work.
(For full details of the experiments, see Protocols.)
1. General Action of Tutin on Mammals.
(a.) Action on Cats.
The effect of various doses upon cats will be shown first.
Effects of a large dose: 9.8 mlgm. tutin (3 mlgm. per kilo body-weight) were injected under the skin of a full-grown female cat, weighing 3.28 kilograms (Exp. 1). The animal was placed in a hutch, and watched continuously. Four minutes after the injection it was noticed that respiration was very rapid (48 to the quarter-minute). The animal seemed sleepy and dazed,
and curled itself down as if to sleep, but at once raised its head, opened its mouth, and panted for breath, like a dog on a hot day. It moved its head slowly from side to side in a dazed, stupid way. Seven minutes after the injection, thick, ropy saliva began to pour from the open mouth, the respirations were even more rapid, and were now audible, almost stertorous, and the whole body was shaken with the force of the respiratory effort. The pupils at this stage were moderate. Ten minutes after the injection the animal got up and began to walk cautiously about the hutch, picking its way like a cat crossing a muddy street. Half a minute later it was suddenly seized with a violent convulsion, which in every respect resembled a typical epileptic fit. The animal fell on its side, and all the muscles of the body were thrown into intense tonic spasm. The head was bent firmly backwards, the back hollowed, the fore and hind limbs rigidly extended, the digits widely separated, the claws extruded. The hairs of the tail became erect, urine and faeces were shot out with considerable violence, the pupils were widely dilated, respiration entirely ceased, and the nose became cyanosed. This tonic spasm lasted thirty seconds, and was followed by clonic spasms affecting the neck, jaw, limbs, and respiratory muscles, which lasted twenty-five seconds more. As the fit passed off, the pupils contracted, the respirations became deeper and slower, and the nose recovered its normal appearance. An interval of five seconds was followed by another fit like the firsts, but the tonic stage did not last so long, and the clonic spasms were more marked. Fit succeeded fit in rapid succession. There were no voluntary cries, but the violence of each seizure caused a choking noise as the air was driven through the glottis. Between the fits the pupils always contracted, and they began to dilate just before each convulsion occurred; during the fit they were dilated to the widest possible extent. This alternation happened so invariably that a commencing dilatation of the pupil could be taken as an indication that a convulsion was imminent. As the animal became exhausted the fits diminished in severity and frequency, the respirations became irregular, infrequent, and gasping, and finally death occurred thirty-one minutes after the injection. From the first the animal gave no indication that it suffered any pain, and from the onset of the convulsions it was unconscious, without either ear or conjunctival reflex. The temperature at death taken in the rectum was 102.4° Fahr. On post mortem examination the right horn of the uterus was found to contain a nearly full-sized foetus, which looked as if it had also been affected with convulsions; one hind leg was twisted over the other, the right forepaw was behind the right ear, and the claws were extruded. Beyond some small haemorrhages in the lungs and a marked congestion of the brain and cord there was nothing noteworthy.
In the next experiment (Exp. 2) the dose was reduced to 2 mlgm. per kilo body-weight, 7 mlgm. of tutin being injected under the skin of a cat weighing 3.5 kilograms. The first symptom noticed (ten minutes after the injection) was trembling of the head and fore part of the body. This was followed, fourteen minutes after the injection, by rapid breathing (56 to the quarter-minute) and by salivation. At twenty-one minutes, slight twitching of the eyelids and ears was noticed. At twenty-four minutes the animal defsecated, discharging a large quantity of faeces. At twenty-five minutes the twitching, which had been gradually getting more marked and more extensive, was severe. At each attack the pupils dilated, but returned to the normal size when the twitching ceased. The respirations were irregular, exaggerated, and suggestive of the Cheyne-Stokes type.
At twenty-nine minutes the animal was seized with a general convulsion, the tonic stage lasting thirty seconds. From this point the convulsive movements continued almost without intermission until the end. The clonic spasms were the more in evidence, but every now and then a tonic seizure would arrest the movements for a few seconds. Gradually the movements became more and more feeble, the respiration slow, irregular, and gasping, and finally the animal died in a tonic spasm fifty-one minutes after the injection had been given. In this case the symptoms appeared more gradually, and the cerebrum seemed less affected than in the first cat. The animal was less dazed and stupid, and frequently “miaued” in a plaintive way, especially after the twitching began. The twitching was a marked feature; it began in a small way, affecting only the eyelids and ears, but the attacks increased in frequency and in severity, and gradually more and more muscles became involved. In the earlier stages, when only the face and neck muscles were affected, the cat at each attack presented the appearance it might have done had it been held and a rapid series of electric sparks discharged close to its face. Later, when the muscles of the shoulders and fore legs were involved, the twitching caused little springs into the air and down again with the fore part of the body, suggesting the appearance of a puppy pouncing at play. When the twitching had got the length of involving the shoulders, it very soon took the form of a general convulsion, and in this case, once convulsions had set in, the symptoms which followed were much the same as seen in the first cat.
In the next experiment (Exp. 3) the dose was considerably reduced, 1.7 mlgm. being injected under the skin of a cat weighing 2.394 kilograms. This is equivalent to 0.75 mlgm. per kilo body-weight. Nothing was noticed for half an hour, and in that time the cat looked quite normal. It then began to swallow rather frequently, as if swallowing saliva, and then suddenly got up and defaecated. It then began to breathe rapidly, saliva could be seen dripping from the mouth, and it vomited. It again attempted to empty the bowels, and now looked miserable, and kept up a constant complaint, though if spoken to it would come forward and purr. Twitching of the face did not appear till thirty-five minutes after the injection, and the first convulsion occurred at forty-one minutes. It lay on its side for a few minutes after the convulsion, and then got up and walked about the cage. Its condition now was much improved on what it was before the convulsion occurred. The breathing was easy, salivation seemed to have ceased, and the twitching which had so annoyed and alarmed the animal was not noticed; but the improvement was not for long. In about a quarter of an hour it began to breathe quickly again, and the twitching returned, and gradually got more and more severe till it culminated in a general convulsion at seventy minutes after the injection. This was a most severe tonic spasm, which lasted four minutes and a half by the watch, and during that time the animal was not seen or heard to draw a breath. From this onwards convulsions continued with intervals of only a few seconds till death occurred, one hour and forty-nine minutes after the injection. In this cat the cerebrum seemed less affected than was the case with the other two. It continued alive to its surroundings almost to the end, and “miaued” voluntarily five minutes before death. It never rose after the second seizure, though it once or twice attempted to do so, and was thrown down by a convulsion. In the intervals between the attacks, running or swimming movements were noticed.
In the next animal experimented upon (Exp. 4) the dose was reduced
to 0.375 mlgm. per kilo of body-weight, 1 mlgm. of tutin being injected under the skin of a cat weighing 2.688 kilograms, and, curiously, symptoms made their appearance earlier in this case than in the last. It defaecated and began to breathe quickly within twelve minutes of the injection; salivation was noticed at fourteen minutes, and at twenty-four minutes it was panting with its mouth open, and vomited. Twitching appeared at thirty-two minutes, and the first convulsion occurred at fifty-five minutes. Convulsions were severe and frequently repeated, and it was thought that the cat would die. It had a severe convulsion at 6.5 p.m., and it was not seen again till 7.15 p.m. It then appeared rather tremulous, and was easily startled, but presented no further symptoms, and was quite well next day. It would seem that 0.375 mlgm. per kilo is very near the minimum lethal dose.
In the last three cats the constant symptoms were defaecation, rapid breathing, salivation, twitching, and general convulsions, and these generally made their appearance in the order named. The first cat did not defaecate voluntarily, and neither of the first two vomited. The vomiting in the last two occurred only once in each case. In the first convulsion in each case the tonic stage was the more pronounced, and was invariably of the opisthotonic type. Later, clonic spasms were more in evidence, and with lethal doses, when the case was making towards a termination, movement was hardly absent for a moment. With the larger doses the effect upon the mental activities of the animals was very marked. From the first they seemed dazed, and once general convulsions had set in they were oblivious of everything. With the smaller doses the cerebrum was little affected, and often after the most severe and prolonged convulsions, in the case of the cat that recovered, the animal would rise and behave as if little had happened, answering when spoken to, and even purring.
(b.) Action on Rabbits.
Rabbits are less readily affected by the poison. The largest hypodermic dose recovered from was 2 mlgm. per kilo (Exp. 5), (in a cat, 0.75 mlgm. per kilo proved fatal). A dose of 2.5 mlgm. per kilo was fatal in two hours and a half (Exp. 6), and doses larger than this killed rather rapidly (Exps. 7, 139). By oral administration a much larger dose than this is required to kill—e.g., in Exp. 8, 7.5 mlgm. per kilo proved fatal in two hours; in Exp. 9, 6 mlgm. per kilo was fatal in twelve hours; while in Exp. 10, 5 mlgm. per kilo was recovered from. In one case (Exp. 161) death preceded by typical symptoms followed the instillation of four drops of a 0.5-per-cent. solution into the conjunctival sac.
Attempts to poison rabbits with fresh tutu-leaves failed, for the animals, though deprived of all other food for several days, steadfastly refused to eat.
Symptoms.—-No important symptoms appear that have not been mentioned as occurring in cats. After a lethal dose the animal at first appears dazed, tends to assume unnatural attitudes—e.g., lies on the abdomen, with the legs projecting in front and behind—and the gait is altered. Respiration soon becomes very rapid, and there may be salivation, though it is not so marked a symptom as it is in cats. Alteration in the size of the pupil is not so noticeable. Twitching of the eyelids, lips, ears, and fore-paws occurs, and is followed by general convulsions. In the convulsion the tonic spasms are not so evident as they are in cats, but they do occur, and are of the opisthotonic type. As a rule, after the first violent convulsive movements are over, the animal continues lying on its side, and
shows almost constant movement, either clonic spasms or running-movements, chiefly of the fore paws, until exhausted. Urination occurs during the convulsions, but, of course, vomiting was not observed. The animal utters no cry, and indicates in no way that it suffers pain. In the later stages it is comatose, and usually dies from exhaustion.
(c.) Action on Guinea-pigs.
No series of experiments was made to determine the minimum, lethal dose in these animals, but it would appear to be rather smaller than, in rabbits. A dose of 2 mlgm. per kilo caused, symptoms in thirty minutes and death in seventy minutes in one case (Exp. 11), while a dose of 1 mlgm. per kilo caused convulsions, but the animal recovered (Exp. 167).
A young guinea-pig was killed by a dose of 1 mlgm. per kilo, while another of the same age (five days) showed symptoms with 0.75 mlgm. per kilo, but recovered. It would appear from this that the young guinea-pig is more easily affected than the adult.
By oral administration a dose of 1.5 mlgm. per kilo was insufficient to produce obvious symptoms, but a dose of 2 mlgm. per kilo repeated in forty-eight hours caused death (Exp. 13).
The symptoms are very like those shown by rabbits—viz., unnatural attitudes, rapid breathing, twitching, general convulsions, and running movements of the limbs. The spasms are more clonic than tonic. When convulsions first appear, the animal tumbles and tosses about in every direction, but soon takes up a position on its side, and then continues in constant movement until exhausted (Exps. 14, 167).
2. Action on Birds.
It is the common opinion that birds are immune to the tutu-poison. It were not strange did a relative immunity exist, for the plump, sweet, attractive-looking berries seem to have been designed by nature for the especial purpose of inducing birds to eat; and that they do eat them freely and without injurious effect is certain. Mr. Maning, author of “Old New Zealand,” quoted by Lauder Lindsay (3), says, “Many kinds of birds live entirely on the tutu-berries when in season… The tui (Prosthemadera novce-zealandice) I have kept tame and fed for months on nothing else.”
Again, birds differ from mammals in having a higher rate of oxidation, a higher temperature, and a peculiar metabolism, which results in the excretion of urates instead of urea in the urine—features which might conceivably have an important bearing on the question of their possible immunity.
Christie (11), as the result of experiment, inclined to the general opinion that birds are immune. He injected in all 40 minims of an ethereal solution of “oil of tutu” into the “chest cavity” of a young rooster. Beyond some slight symptoms which were attributed to the ether, and a marked increase in the frequency of defaecation, no characteristic effect was noted. He observes that “there was no twitching, although the dose (40 m.) is twice as much as is necessary to convulse a cat”; and concluded that the bowels are chiefly affected in birds, and that, therefore, they are saved by rapid excretion by this channel. When the above statement is more closely examined it is found that Christie administered probably not more than 1 mlgm. of pure tutin per kilo. The symptoms were wanting because the dose was inadequate.
Experiments that have been made by the present writer on pigeons prove that birds are not immune to tutin. The minimum lethal dose by mouth, however, was found to be high, and this probably accounts for the apparent immunity, the ingestion of very large quantities of berries being necessary to produce toxic symptoms.
In the first experiment (Exp. 15) it unfortunately happened that the pigeon used displayed a peculiar tolerance to the poison. This misled to the belief that the minimum lethal dose was much higher than it really is. The bird, weighing 314 grams, was subjected to successive doses, equivalent to 2, 4, 6, 8, 10, 12, 16, 20 mlgm. per kilo of body-weight. The solution, of tutin was given by the mouth, and the experiment extended over a period of ten days. The smaller doses had little effect. For a few hours after each dose the bird was dull, apathetic, and disinclined to eat or move about. It stood in a corner of the cage, with its feathers puffed out, blinking heavily, and presented the appearance of a bird that had overeaten itself, and was trying hard to go to sleep, but was too uncomfortable to succeed completely. The last dose (20 mlgm. per kilo) was given on an empty crop, and as no ill effect was anticipated—it being thought at this time that in all probability birds really were immune—it was not closely watched. But forty-five minutes later attention was attracted by the flapping of its wings, and it was found lying on its back in convulsions. One hour after the administration of the poison it was dead. Its weight at death was 345 grams—a gain of 31 grams in ten days, which would seem to indicate that tutin has no injurious influence on general metabolism. That the bird actually received the doses stated is quite certain. The solution was very carefully measured in a hypodermic syringe having a running-nut on the piston-rod, and dropped into the beak, which was held open by the finger. There was no difficulty in the administration, and every particle of the fluid was swallowed.
As 20 mlgm. per kilo was obviously too great a dose, it was decided to give to the next bird I mlgm. per kilo in excess of the largest ineffective dose on the first bird. To this end a dose equivalent to 17 mlgm. per kilo was administered by mouth to a fasting pigeon weighing 319 grams (Exp. 16). In this case toxic symptoms—viz., twitching and tremulousness of the head and wings, and attempts to vomit—made their appearance within two minutes, violent convulsions appeared in four minutes, and death occurred sixteen minutes after the administration.
In contrasting these two experiments, it will be noticed that the “time to kill” was greater in the first case by forty-four minutes, although the dose exceeded the dose in the second case by 3 mlgm. per kilo. This observation, taken in conjunction with the fact that symptoms did not appear in the first case with a dose of 16 mlgm. per kilo, suggested the possibility of the first pigeon having acquired some degree of immunity or tolerance by the poison having been administered in gradually increasing doses over a lengthened period. To test the validity of this supposition, another pigeon was treated in the same way (Exp. 17). The experiment extended over a period of three weeks, and a maximum of 10 mlgm. per kilo was reached without the appearance of any marked symptoms, but 12 mlgm. per kilo proved fatal.
The rapidity with which symptoms ensued after a dose of 17 mlgm. per kilo, and the speed with which a fatal termination was reached, pointed to the dose being well above the lethal minimum. A dose equivalent to 15 mlgm. per kilo was therefore given by mouth to a pigeon weighing
357 grams (Exp. 18). No symptoms followed beyond those that generally appeared in the first experiment after a non-lethal dose had been administered. One hour and three-quarters later the bird was apparently normal and on the day following the experiment was quite well. Here a dose of 15 mlgm. per kilo was without effect. It was noted that the crop in this bird was full, but, though this was recognised as probably influencing the result in some degree, the fact that the first bird was unaffected by a dose of 16 mlgm. per kilo was so striking that it was thought likely that 15 mlgm. per, kilo was below the lethal minimum. It was therefore decided to give a dose equivalent to 16 mlgm. per kilo to another pigeon (Exp. 19). In administering the poison to this bird, by an accident a few drops of the solution were lost and the deficiency was made good by an allowance of 5 extra minims (Exp. 15). In three minutes the bird was in convulsions, and in nine minutes was dead. As no satisfactory conclusion could be drawn from this experiment it was repeated (Exp. 20), but the subject became convulsed within a minute and a half, and died five minutes after the administration.
It was then decided to repeat the dose of 15 mlgm. per kilo, using the bird that had previously withstood this dose, but taking care that its crop should be empty (Exp. 21). At 5.14 the dose was given. No symptoms appeared till seven minutes had elapsed, when it began to retch. In twenty minutes convulsions appeared, and in thirty-five minutes it was dead. It was noted that the onset of symptoms was slower and the time to kill longer in this case than in the case where 16 mlgm. per kilo were given (thirty-five minutes as against five minutes). It was therefore thought worth while to repeat the dose of 15 mlgm. per kilo, using a bird that had not been starved. This was done (Exp. 22), with the result that the bird died in twelve minutes.
It being now clear that the minimum lethal dose was much exceeded, two pigeons were taken (Exps. 5, 23, 24), and to one was given a dose equivalent to 13 mlgm. per kilo, to the other 12 mlgm. per kilo. In the case of the bird with the larger dose, death resulted in nineteen minutes; but the bird that had received 12 mlgm. per kilo showed a very gradual onset of symptoms, and did not die till two hours and sixteen minutes after the administration. This bird was a young one (still squeaking), though it weighed 363 grams. It was thought that the age in this case may have influenced the result, so a dose of 12 mlgm. per kilo was given to an adult bird. Death occurred, however, in forty-six minutes—a hundred minutes earlier than in the case of the young bird (Exp. 25).
A drop in dosage was now made to 9 mlgm. per kilo, and the bird that received this dose was not affected (Exp. 26). The same result was noted in a bird receiving 9.5 mlgm. per kilo (Expl 27); but the next bird, which received a dose of 10 mlgm. per kilo, died in forty-five minutes (Exp. 28). This was a young bird (squeaking), weighing 330 grams. The same dose (10 mlgm. per kilo) was therefore given to an adult bird (Exp. 29), which recovered after exhibiting symptoms such as vomiting and slight convulsive movements of the wings.
Of the four birds that had received 10 mlgm. per kilo, one only (a young bird) had died. A dose of 10.25 mlgm. per kilo was therefore given, to, an adult bird, with the result that it died in seventy-five minutes. (Exp. 30).
The results of these experiments are tabulated on the following page (Table 1).
|Exp.||No.||Weight.||Mlgm. per Kilo||Crop.||Age.||Result.||Time to kill.|
|12, 16, 20||"||"|
|21||6||345||15||Empty||"||Died (same bird as No. 5)||351/2|
|17||11||370||1.5, 2, 4, 7, 9, 9.5, 10, 12||Half-full||"||"||(?)|
It will be seen that the highest dose recovered from was 10 mlgm. per kilo, and the lowest dose that killed was 10.25 mlgm. per kilo. It may be taken as proved, then, that birds are not really immune, as has been supposed, but they are able to withstand a very high dose of the poison by oral administration.
In noting the current opinion that birds are immune, Easterfield and Aston state (7) that cases have come under their notice in which domestic fowls have been poisoned by eating tutu-berries. This seems barely credible. The lethal dose by the mouth in birds is so high, the percentage of tutin in the seeds so low, the size of the berry in proportion to the size and number of the seeds it contains so great, that it may be doubted if the crop of the ordinary fowl could comfortably accommodate the large number of berries that would be required to provide a lethal dose of tutin. For example Easterfield and Aston found (7) that the dried seeds of C. ruscifolia contained 22.8 per cent. of a green oil, of which 0.18 gram administered to a small kitten produced only very mild symptoms of poisoning. Assuming that this 0.18 gram contained 0.0003 gram tutin—a dose which the present writer has found to be feebly toxic to a cat (Exp. 4)—the amount of tutin in the dried seed must be about 0.038 gram in 100 grams. The minimum lethal dose by the mouth in birds is just over 10 mlgm. per kilo, so that in order to get a fatal result, a bird weighing 0.3 kilo (the average weight of a pigeon) would require to eat a little more than 8 grams dried seed, or 150 grams—over 5 oz.—of the fresh fruit. (This statement is based on the relative weights of seeds and berries.) An average-sized domestic fowl would require about 1 lb. of the fruit, and that, of course, assuming that all the tutin would be extracted from all the seeds. To throw light on this point, 5 grams (all that was at hand) of cleaned dried seeds bound with a little moist flour were fed into the crop of a pigeon weighing 314 grams (Exp. 31). It displayed
no symptoms, was quite well the next day, and continued well. Apparently unchanged seeds were recovered from the faeces.
An attempt was made to induce a domestic fowl to eat some of the fresh fruit (Exp. 32). After being deprived of food for two full days, it was offered a heaped plateful (5 oz.) of berries. After eating about 1 oz. to 1 ½ oz. it desisted, and ate no more, although it had no other food given it for two days longer. It displayed no symptoms.
The immunity of birds from poisoning by the seeds under natural conditions may be explained perhaps by—(1) The large number of seeds that must be ingested in order to provide a lethal dose; (2) the seeds being small and hard, and probably passing through the animal unchanged, as was noted to be the case in Exp. 31. Their relative immunity may possibly be accounted for in some measure by—(1) The higher rate of oxidation; (2) the fact that the kidneys in birds are able to secrete substances in a semi-solid condition, certain normal constituents of the urine—urates and uric acid—having been observed by Bowman (25) in the cells of the tubules. This would suggest that the excretory power of the tubule cell for insoluble substances is greater in birds than in mammals. If this suggestion be applied to the case of a poison that is with difficulty soluble in water circulating in the blood, it may be conceived that this special power of the tubule cell in birds will allow of a more rapid elimination of the toxic body, and so confer a relative immunity. This, of course, applies where the poison is gradually received into the blood, as when absorbed from the alimentary tract. Where, on the other hand, the toxic body is rapidly introduced into the blood, as when given hypodermically, the cells of the tubules have not an opportunity of exercising their special power, and so the relative immunity is not so marked, or does not obtain. This may be illustrated by Exp. 53, where a dose of 5 mlgm. per kilo was given hypodermically to a pigeon weighing 335 grams, with the result that convulsions appeared in twenty minutes, and continued without intermission till death occurred, forty-two minutes after the injection.
The question of the relation of the liver to a glucoside like tutin is an interesting one, and especially so in this connection. In birds the veins from the crop are branches of the jugulars, and therefore a toxic body absorbed from the crop, as must have been the case in the experiments cited, enters the circulation direct. In mammals, on the other hand, the gastric vein discharges the blood from the stomach—which, so far as the absorption of poisons is concerned, corresponds to the avian crop—into the portal vein, and therefore the toxic body must pass through the liver before it enters the general circulation. Has the liver-substance any power to increase or decrease the toxicity of tutin? To determine this, experiments were made on fish. The result was doubtful, but the indications are that the liver-substance does increase the toxic power, possibly by separating the glucose part of the compound from the poisonous part, and so freeing the latter of an innocuous encumbrance.
Symptoms in Birds.—With a massive dose the animal is almost immediately, with hardly any premonitory symptoms, seized with general, convulsions. The wings are rigidly extended to the full, and are flapped violently. The head is retracted till it lies firmly pressed against the back between the wings, and the bird is thrown over backwards. It turns over and over, and flops about in every direction until exhausted. This period of violent movement does not last more than a few minutes. A final somersault lands the bird on its back, and it lies, with its head drawn back beneath its body,
in continuous movement—flapping its wings and clawing at space with its feet. Respiration is spasmodic, and the forcible expulsion of air from the lungs is distinctly audible. The pupils are widely dilated—no, iris being visible—and the bird is in a state of coma, oblivious to everything, and apparently suffering no pain. In the later stages the head is brought forward, the eyes are kept closed, and the movements, which have continued without intermission from the onset, gradually become more and more feeble, and finally cease at death almost imperceptibly. The legs continue in movement longer than the wings.
With a smaller though still lethal dose the first change is a heaviness and drowsiness, giving the bird a peculiar sleepy, stupid look. The eyes are blinked heavily, and seem to be kept open only with the greatest difficulty. This is succeeded by attempts to vomit, and, if the crop contains anything, by actual vomiting. Then follow tremulousness of the head and wings and sharp spasmodic blinking of the eyes. The head is frequently drawn back sharply, and one or other or both wings are involuntarily extended for a moment. These seizures of the head and wings recur at frequent intervals, and gradually become more severe, making it difficult for the bird to keep its feet. In the more severe seizures it is thrown back on to its tail, and sits there for a moment until the clonic movements of the extended wings have ceased. At last the bird is thrown over on to its back, and, with continuous convulsive movement, the case hastens to a termination.
With non-lethal doses little is to be observed beyond the initial drowsiness and tremulousness, and perhaps vomiting and slight convulsive movements of the wings. In none of the experiments did any of the birds recover that reached the stage of being thrown over on to the back.
In none of the birds experimented upon was Christie's observation repeated that there is increased frequency of defaecation.
3. Action on Reptiles.
The only animals of this class available for experiment were three small lizards of the species Lygosoma moco, the common lizard of New Zealand. They are very small animals, these specimens weighing 4, 5, and 7 grams, and are not very suitable for experiment.
(Exp. 33.) The first lizard (4 grams) was given a dose 5 mlgm. per kilo by hypodermic injection, and it became convulsed, and died in about two hours.
The second lizard (7 grams) (Exp. 34) received 4 mlgm. per kilo hypodermically, and died four hours and three-quarters later, after showing severe and oft-repeated convulsions. Opisthotonos was well marked, the animal bending backwards till head and tail met. There were also very definite clonic spasms of the limbs. For over an hour the animal was in almost constant movement, contorting itself and twisting in every direction.
(Exp. 35.) To the third lizard was given a dose of 3 mlgm. per kilo, but beyond exaggerated respiratory movements it displayed no symptoms, and was quite normal on the day following the injection.
Lizards are therefore affected in the same way as other animals. The lethal dose is between 3 and 4 mlgm. per kilogram.
4. Action on Amphibia.
The frogs experimented upon belong to the species Hyla aurea. This is not the native frog of New Zealand, but has been introduced from Aus-
tralia, and is now the common frog. They were kept till wanted in a large wooden box, in a cool, dark cellar, and supplied with water and fresh grass-sods, but no special provision was made for feeding them. Under these conditions they lived well, and remained healthy.
Symptoms (Exps. 54 to 58 inclusive).—With a lethal dose, the first symptom usually noticed was increased frequency of the respiratory movements. To this succeeded lethargy and muscular enfeeblement, the animal crawling in a laboured way, and trailing its hind limbs. If touched now it would attempt to hop, but made little progress from inability to draw the hind limbs completely beneath the body. If left alone it would lie prone on the belly with the limbs extended, and make no effort to move. When convulsions occur, as they usually do, though they are not such a marked feature as in mammals, tonic spasms are as a rule more apparent than clonic, and the hind limbs more affected than the fore limbs. Often the animal raises itself rigidly to the full extent of the fore limbs, and open its mouth spasmodically two or three times in succession. If handled much, or compelled to undergo severe muscular exertion, such as struggling violently to recover the normal position when placed on the back, convulsions appear at once. This recalls the fact that a “tooted” sheep often displays no very obvious symptoms until it is “worked.”
A series of experiments was made to determine the minimum lethal dose (Exps. 36 to 52 inclusive), and the results are presented in tabular form below (Table II). In the early experiments the animals, after receiving the injection, were placed under bell-jars on the laboratory-table. Here they were deprived of moisture, and exposed to a strong light. Under these experimental conditions the results were confusing, frogs with larger doses displaying no symptoms, while animals with lesser doses died. The method was therefore improved upon. Each frog after receiving its injection was placed in a small box containing a moist grass-sod in a cool, dark cellar, and a striking uniformity in result was at once obtained.
|36||27.5 grams||1mlgm. par kilo||Died||Unfavourable surroundings.|
|42||31"||4"||"||Under improved conditions.|
Table II shows that the highest non-lethal dose attained was 10 mlgm. per kilo. Doses of 10.25 mlgm. and over were invariably fatal. Of the frogs
that received 10 mlgm. per kilo, one died, and both the others showed convulsions and were very ill for more than twenty-four hours. Severe symptoms were also present with 9 mlgm. per kilo. It was noticed that frogs experimented upon in the winter were more susceptible to the poison than those used in the summer.
The relatively high lethal dose of tutin in frogs may have some connection with their mode of respiration. To a large extent this is cutaneous, and therefore a drug such as tutin, which owes its lethal power largely to its influence on the respiratory centre, might a priori be expected to be less lethal to these animals than to those that have only a lung-respiration. The fact that lizards, which have a dry skin and presumably only a slight cutaneous respiration, succumb to about one-third the lethal dose for frogs points in this direction.
Even with large doses the course of events in frogs is comparatively slow. For example, in Exp. 54, where about 60 mlgm. per kilo was given, the animal lived for an hour and a half.
5. Action on Fishes.
(a.) General Action.
There is an advantage in using small fishes to test the pharmacological action of drugs, because large numbers can be dealt with, and thus the factor of idiosyncrasy, which so often confuses results, is in great measure eliminated. Moreover, fishes can be placed in a watery solution of the drug, and the symptoms and time taken to kill easily noted. Recently Sollmann (26) has used funduli and sticklebacks in this way, and suggests that the method might be used to study the antagonism of drugs.
In the first experiments of this kind undertaken by the present writer young trout procured from a fish-hatchery were used. The fry were kept till required in a trough with an overflow, placed beneath a running tap, and were fed on fresh minced liver. The experiments were carried out in the following way: 60 or 100 c.c. of tap-water, to which a solution of tutin had been added in definite quantity, were placed in a small wide beaker, and the fish transferred to it. Several beakers containing various strengths of tutin, and one beaker containing tap-water only, as a control, were set on together, and watched at intervals during the day.
Symptoms.—The young trout were about 1 in. to 1 ½ in. long, and in good condition. When first placed in the test-beakers they became very excited if the tutin solution were strong. After a period of excitement, during which rotatory movements on the long axis of the body were frequently seen, they became quieter, and then it was noted that the gill-movements were much exaggerated. After a time they lost their power of maintaining equilibrium, and lay on one side gasping. From this position they would frequently spring up and swim round the vessel, still on the side, and then resume their position on the bottom. Shortly before death they often turned belly upwards.
Minimum Lethal Dose.—In order to determine the minimum lethal dose, solutions of different strength were tried, and the results are given in the next table (Table III).
In these experiments the quantity of fluid used was small. Death frequently occurred in the control when the experiment lasted over twelve hours. But the results show that the fatal effects begin when the percentage is about 0.001, and are very definite at 0.004 per cent. This corresponds
to a dose of 10 to 40 mlgm. per kilo. For example, if we consider that a dose of tutin—say, 10 mlgm. per kilo—injected under the skin of a mammal becomes rapidly and equally diffused through all parts of the body, its concentration at any point would be 0.01 gram (10 mlgm.) per thousand. In the case of a fish floating in a solution of tutin, the amount of the fluid is so much larger than the fish that the diffusion of the tutin into its body will not materially reduce the concentration of the poison in the fluid, and so the tissues of the fish will be subjected to the action of 0.001 per cent., or 10. mlgm. per kilo. The fatal dose in fish may therefore be said to be about 40 mlgm. per kilo under these experimental conditions.
|Exp. No.||Percentage of Tutin in Water.||Volume.||Number of Fish.||Time necessary to kill.|
|59||10 drops of saturated solution in 100 c.c. (about 0.1 per cent.)||100||One||About 43 minutes.|
|60||0.125 per cent||60||"||19 minutes.|
|61||0.0625 "||60||"||27 "|
|62||0.03125 "||60||"||50 "|
|63||0.0156 "||60||"||40 "|
|64||0.0078 "||60||"||56 "|
|65||0 004 "||60||"||111 "|
|66||0.001 "||50||"||4 ½-19 hours.|
|67||0.00075 "||50||"||4 ½-19 "|
|68||0.0005 "||50||"||Alive 15 hours 48 min. later.|
|70||0.03125 "||50||"||50 minutes.|
|71||0 004 "||50||"||About 8 ½ hours.|
|72||0 003 "||50||"||"|
|73||0 002 "||50||"||Between 8 ½ and 18 ⅓ hours.|
|74||0.001 "||50||"||19 ¾ hours.|
|75||0.00075 "||50||"||About 30 minutes.|
|76||0 0005 "||50||Two||One died between 8 ½ and 18 ⅓ hours; the other lived, and was alive and well four days later.|
It was intended to use trout-fry in experiments that were made to compare the action of tutin with that of other members of the same group, and to test the effect of remedies; but the supply suddenly failed, so recourse was had to another small fish, known locally as the minnow, which abounds at certain seasons of the year in the waters of the Otago Harbour and Lake Logan. This fish (Galaxias attenuatus) is peculiar to the South Island of New Zealand, Tasmania, and Tierra del Fuego (33). It measures 1—3 ½ in. long, is equally at home in brackish and fresh water, and will live in a trough in the laboratory for weeks if fed on liver and supplied with fresh running water. It is semi-transparent, and the heart can be seen beating. After a great many experiments it was found that the best results were obtained when three fish were used, and placed in a fairly wide (7-in.-diameter) enamelled bowl, containing 1,000 c.c. fluid. It was found necessary to almost completely cover the top of the bowl with a piece of wood, for if left uncovered the fish were apt to leap out. This happened several times, and was often more aggravating to the experimenter than
disastrous to the fish, for if discovered within an hour or so, wiped clean, and returned to the water it would recover completely. Three of these fish will live in 1,000 c.c. of water from thirty-six to forty-eight hours or more, and are apparently quite comfortable, even when the water is much fouled with their own excreta.
Sollmann states that 0.001 per cent, picrotoxin is fatal to fundulus in five to nine hours. His specimens (5 cm.) were not much smaller than the minnows used in the experiments recorded here. He used only 150 c.c. of water to each fish, and admits that occasionally one of the controls would die. It is probable that the amount of water used by Sollman was too small, for in the experiments made by the present writer to compare the effects of tutin and picrotoxin, it appeared that in the early experiments, where a small quantity of water (200 c.c.) was used, death occurred in four hours when the concentration was 0.001; whereas in the later experiments, with three fish in 1,000 c.c. fluid, 0.001 was fatal to only one fish of the three in twenty-four hours. This shows the importance, when using fish in this way, of allowing a sufficient quantity of water.
Symptoms.—The symptoms shown by the minnows when three were put together in 1,000 c.c. of a lethal percentage (0.005) of tutin were as follows: After two to four hours, with no symptoms, the fish began to swim near the surface of the fluid, and became excited in their movements, swimming vigorously about, and even leaping repeatedly out of the water. At this stage they were frequently observed to emit bubbles of gas from the mouth. This discharge of gas, and the inability to sink, point to a derangement of the function of the swim-bladder. Later they lose their power of maintaining the normal position, and swim about near the surface, turned on the side. Now and again they recover the upright position, and swim excitedly about for a minute or two, and then fall back on the side. Later on they lie at an angle with the surface of the fluid, often with the head down, and swim about feebly in this position; and later still they lie on the side on the bottom of the dish, and the gill-movements are laboured. At intervals they spring up from this position, and swim round on the side, while appearances suggesting convulsions are seen—viz., bending of the trunk, accompanied by shuddering movements. At death it was invariably noticed that the gill-movements ceased before the heart stopped beating. (For protocol of symptoms, see Exp. 108.)
Table IV shows that under the above-mentioned experimental conditions a percentage of 0.001 was necessary to cause symptoms to appear, but was not necessarily fatal unless other deleterious conditions, such as too small an amount of fluid, were present (e.g., Exps. 77, 78, 79 in Table IV show fatal results within six hours, but under improved conditions 0.0035 was fatal to only one fish out of three); while a percentage of 0.005 caused symptoms within a few hours, and was generally fatal to all three fish within twenty - four hours. Between these limits various results were obtained: thus in 0.0035 per cent. two of the three fish recovered and one died, while in 0.003 per cent. and in 0.004 per cent. two of the three died and one recovered. But in these experiments, Nos. 89, 91, and 95, tin dishes were used, and the water became rusty before the end of the experiments.
In order to insure a lethal effect, the dose was therefore raised to 0.005 per cent., and experiments were made to test the effect of various reagents in increasing or diminishing the toxic power of tutin. The effect of tutin was also compared with that of picrotoxin.
|Exp No||Percentage of Tutin used:||Volume of Fluid.||Number of Fishes.||Time of Onset of Symptoms.||Result.||Time of Death.|
|83||0.001||1,000||Three—(a)||Slight symptoms at 9 ½ hours||Not fatal|
|(b) and (c)||No symptoms||"|
|86||0.0015||1,000||Three—(a)||1 hour||Fatal||1 5/6|
|(b) and (c)||No symptoms||Not fatal|
|88||0.00175||1,000||Three—(a)||4 ½ hours||Fatal||4 5/6|
|(b) and (c)||No symptoms||Not fatal|
|(a)||2 ¾ hours||Fatal to all||7|
|89||0.003||1,000||Three(b)||9 ½ "||12|
|(a)||4 "||Fatal||4 ½|
|91||0.0035||1,000||Three (b)||Slight symptoms from 7 to 10 hours||Not fatal|
|(a)||3 3/11 hours||Fatal||12|
|95||0.004||1,000||Three(b)||8 "||"||8 ½|
|(c)||Slight symptoms at 12 hours||Not fatal|
|(a)||7 ½ hours||Fatal to all||12-22|
|106||0.005||1,000||Three(b)||7 ½ "||12-22|
|(c)||6 ½ "||24|
|(a)||2 ⅘ hours||Fatal||12 ½-25|
|108||0.005||1,000||Three(b)||4 ⅓ "||"||12|
|(106 used a second time)||(c)||5 ½ "||Not fatal|
(b.) Comparison of Action of Tutin and Picrotoxin.
Sollmann's statement that 0.001 per cent. picrotoxin causes death in four to nine hours was tested by the improved method (see Table V, No. 85). Three fish were put into 1,000 c.c. of 0.001 picrotoxin at 11 a.m. Up to 8.30 p.m. no symptoms were seen, though the fish were under observation all day. At 8.30 they began to swim more frequently near the surface, and when seen next day one was dead, but the others were unaffected, and were returned to the trough at the end of twenty-four hours. A 0.001 per cent. of tutin (Exp. 83) showed almost similar results. Previous to this an experiment where 0.00075 per cent. and 0.0005 per cent. picrotoxin and the same strengths of tutin were tested gave doubtful results, because the amount of fluid was too small (Table V, Nos. 81, 82, 78, 79).
In other experiments picrotoxin was compared with tutin in a strength of 0.004 per cent. All the picrotoxin fish died—two at twelve hours and one at 29 ½ hours (Exp. 100); while of the tutin fish two died—at twelve hours and at eight hours and a half—and one recovered (Exp. 95). With a 0.005-per-cent. solution of picrotoxin all three fish died—viz., at 3 ½ hours, at 11 ¾ hours, and at 12 ⅔ hours (Exp. 107); while tutin of the same strength caused death to all at 22 ¼ hours and at twenty-three hours (Exp. 106): so it would appear that picrotoxin is more lethal than tutin in equal percentages.
|Exp No.||Percentage of Drug.||Volume of Fluid.||Number of Fisbes.||Time of Onset of Symptoms.||Result.||Time of Death.|
|80||Picrotoxin, 0.001||200||One||Not observed||Fatal||4|
|81||Picrotoxin 0.00075||200||"||5 hours||"||6|
|77||Tatin, 0.001||200||"||Not observed||"||4 ½|
|78||Tutin, 0.00075||200||"||5hours||"||6 ⅓|
Control to the above fishes under the same conditions (200 c.c. fluid) showed symptoms of asphyxiation in four hours.
|Exp. No.||Percentage of Drug.||Volume of Fluid.||Number of Fishes.||Time of Onset of Symptoms.||Result.||Time of Death.|
|85||Picrotoxin, 0.001||1,000||Three||9 ½ hours||Fatal to one||22 (about).|
|83||Tutin, 0.001||1,000||"||9 ½ "||All recovered|
|(a)||3 ½ "||Fatal to all||29 ½|
|(b)||3 ½ "||12|
|(c)||6 ½ "||12|
|(a)||3 ¾ "||Fatal||12|
|(b)||8 "||"||8 ½|
|(c)||Slight symptoms at 12 hours||Recovered|
|(a)||1 ⅔ hours||Fatal to all||3 ½|
|106||Tutin, 0 005||1,000||Three—|
|(a)||7 ½ hours||Fatal to all||22 ¼|
|(b)||6 ½ "||23|
|(c)||7 ½ "||22 ¼|
|(a)||1 ½ "||Fatal to all||3|
|(b)||1 1/12 "||6 ½|
|(c)||5/6 "||2 ¼|
It was then resolved to compare picrotoxin with tutin in equimolecular solutions. The molecular weight of tutin (C17 H20 O7) is 336, and that of picrotoxin (C45 H50 O19) is 887; hence equal weights of tutin and picrotoxin contain by no means the same number of molecules. Accordingly two solutions were prepared, one of 0.005 per cent. tutin and one of 0.0132 per cent. picrotoxin (0.005 × 887/336), and the experiment was carried out as before, three fish (one large, one medium, and one small) being added to each bowl (Exps. 105, 106). The fish in the picrotoxin solution began to show symptoms first. The small fish came to the surface after fifty minutes, and in sixty-five minutes it was on its side. The medium fish also was much excited. In an hour and a half the large fish was found swimming
near the surface and emitting air-bells, while at this time the other two fish showed quivering movements of the trunk. In two hours and three-quarters the small fish was dead. In three hours the larger one was dead, and the medium-sized one died in five hours. So that 0.0132 per cent. picrotoxin killed all the fish in five hours. Meanwhile the fish in the equri-molecular solution of tutin began to show symptoms at the end of three hours and three-quarters, which continued till late at night—twelve hours after the experiment was begun. The next morning two were found dead (between twelve and twenty-two hours after the experiment was begun), and the last one (medium-sized) died at twenty-four hours (Exp. 106). So the result shows that in equimolecular solutions picrotoxin is more lethal than tutin.
One must be cautious, however, in applying these results to the case of mammals. In fishes the swim-bladder regulation is disturbed both by picrotoxin and by tutin, and of the two, picrotoxin seems to have the greater influence, for air-bells are more frequently seen to be emitted by fish subjected to the action of this drug. The swim-bladder—of such paramount importance in the case of fishes—has no analogue in mammals, and it is therefore quite probable that the lethal power of these poisons may be reversed in this class of animals. Indeed, the experiments on cats (Exps. 4, 168, 169, 171) justify this statement.
(c.) Effect on Fishes of Tutin Solution that has been hydrolysed.
It has been a constant observation in the case of both lower animals and of human beings that tutu poisoning is more lethal when the stomach is empty. This suggested the possibility of the toxicity of the tutin being increased by the action of the hydrochloric acid in the stomach. Experiments on fishes were therefore undertaken to test this. To 8 or 10 c.c. of a 0.5-per-cent. solution of tutin an equal amount of 0.4 per cent. hydrochloric acid was added, and the mixed fluid kept at a temperature of 37° C. for different lengths of time in different experiments. It was then exactly neutralised with a solution of soda of corresponding strength, and made up to 1,000 c.c.
In the first experiment (Exp. 92) of this kind 7 c.c. of tutin solution was so treated (0.0035 per cent. tutin), and a control (Exp. 91) of a solution of untreated tutin in corresponding strength was used. In the hydrolysed tutin solution one fish showed symptoms at three hours and three-quarters, which continued till it died, at seven hours and a half. Another began to show symptoms at eight hours and one-third, and died between ten and twenty-two hours; while the third showed symptoms at nine hours, and died at 22 ½ hours. In the control of untreated solution one fish showed symptoms at four hours and died at four hours and a half, and the other two, of which one showed symptoms from the seventh to the tenth hour, recovered.
Another experiment of the same kind was then made, a slightly higher percentage of tutin being used—viz., 0.004 instead of 0.0035. Two testtubes were taken, each containing 8 c.c. of a 0.5 per cent. solution of tutin. To one, A (Exp. 94), 8 c.c. of 0.4 per cent. HCl were added; so that the total percentage of HCl in the fluid was 0.2. This mixture was kept at 37° C. for one hour, then neutralised and made up to 1,000 c.c. To the 8 c.c. tutin solution in the other test-tube B (Exp. 95) the same amount of HCl was added, but it was immediately neutralised, and then kept at 37° C. for one hour, and made up to 1,000 c.c. In each of these two solutions
three fish were placed, and they were observed at frequent intervals during the day. In B (control), symptoms began in three hours and three-quarters in a fish which died in twelve hours; and at eight hours in a fish which died in eight hours and a half. The third fish showed slight symptoms when last seen at night, twelve hours after the experiment was begun, and was apparently quite well the next day. After thirty hours this fish was removed, the fluid filtered, and three other fish placed in the solution. This second group of fishes was affected as follows: When last seen at night; two hours after the experiment was begun, they seemed unduly excited, and next morning all three were dead—i.e., within sixteen hours of the commencement of the experiment (Exp. 98). (In order to test whether a second use of the solution would influence the result, a control of 1,000 c.c. water that had been used the day before was also filtered, and three fishes placed in it. These fish were quite unaffected at the end of the experiment, so that one may disregard the fact that the control solution mentioned above was used twice.)
|Exp No.||Strength of Tutin.||Hydrolysed or not.||Volume of Fluid.||Number of Fishes.||Onset of Symptoms.||Result||Time of Death.|
|Per Cent.||C c||Hours.|
|92||0.0035||Hydrolysed 30 min. at 37° C.||1,000||Three—|
|(a)||3 ¾ hours||Fatal to all||7 ½|
|(b)||8 ⅓ "||10-22|
|(c)||9 "||22 ¼|
|93||0.0035||Not hydrolysed (control to 92)||1,000||(a)||4"||Fatal||4 ½|
|(b)||Slight symptoms, 7-10 bours||Recovered|
|(c)||No symptoms||Not fatal|
|94||0.004||Hydrolysed 1 hour at 37° C.||1,000||Three—|
|(a)||5 hours||Died||25 ⅓|
|97||0.004||Same fluid as 94, filtered||1,000||(a)||3 "||Died||16|
|(b)||16 "||"||22 ½|
|95||0 004||Control to above||1,000||(a)||3 3/11 hours||Died||12|
|(b)||8 "||"||8 ½|
|98||0.004||Same fluid as 95, filtered||1,000||Three||2 "||Fatal to all||16|
|111||0 005||Hydrolysed 1 hour at 37° C.||1,000||(a)||6 ⅔ "||13 5/6|
|(b)||5 ⅓ "||8 1/6|
|(c)||5 ⅓ "||9 1/6|
|106||0.005||Untreated||1,000||(a)||7 ½"||Fatal to all||22 ¼|
|(b)||6 ½ "||23|
|(c)||7 ½ "||24|
Now, with regard to the hydrolysed tutin (Exp. 94), to which the foregoing served as control experiments: In the first set of three fishes, one showed
symptoms at the end of five hours, and died in 25 ⅓ hours; a second showed symptoms at the end of six hours and a quarter, and died at twenty-eight hours; and the third showed slight symptoms at the end of eight hours, and recovered. The fluid was filtered, and three fresh fishes introduced, and of these one died within sixteen hours, one at 22 ½ hours, and one recovered (Exp. 97).
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A third experiment (Exp. 111) was made with 0.005 per cent. tutin in the same way. Symptoms appeared in two of the fish at five hours and a third; these died at eight hours and a sixth and nine hours and a sixth respectively. The third showed symptoms at six hours and two-thirds, and died at 13 5/6 hours. Usually fish live in a 0.005-per-cent. solution of tutin for an average of seventeen hours.
To summarise the results of these three experiments: Twelve fish were subjected to the influence of hydrolysed tutin (0.0035 per cent. to 0.005 per cent.). Of these, ten died at an average period of fourteen hours; the other two showed symptoms, but recovered. In the corresponding control solutions, of the twelve fish, nine died at an average period of 16.9 hours; two showed symptoms, and recovered; and one showed no symptoms. It would seem to follow that hydrolysed tutin is more toxic than tutin itself; but the rise in toxicity is not great, and in marked contrast to the action which takes place in dilute alkali at corresponding strength.
(d.) Action of Alkalis on Tutin.
Attention having been arrested by the fact that animals poisoned by tutin are frequently reduced to a state of coma, which may possibly contribute towards a fatal termination, the use of alkalis as a remedial agent suggested itself. Hughes had found that lime destroyed the activity of the tutu poison, and Easterfield and Aston were unable to recover tutin after a solution treated with lime had been evaporated to dryness.
|Exp No.||Percentage of Tutin.||Treatment by Alkah||Volume of Fluid.||Number of Fishes.||Time of Onset of Symptoms||Result.||Duration of Experiment.|
|96||0.004||Treated with 0.2 per cent. NaOH at 37° C. for 1 hour||1,000||Three||No symptoms||All lived||30|
|101||0.005||Treated with 0.2 per cent. HaOH at 37° C. for 20 minutes||1,000||"||"||"||25|
|102||0.005||Ditto, for 40 minutes||1,000||"||"||"||25|
|109||0.005||Ditto, for 10 minutes||1,000||"||"||"||30|
|112||0 005||Ditto, for 5 minutes||1,000||"||"||"||26|
The experiments were done in the same way as in the case of HCl: 10 c.c. of a 0.5-per-cent. solution of tutin was rendered alkaline with the solution
used in the foregoing experiments to neutralise the weak hydrochloric acid. The alkalinity used was equivalent to 0.2 per cent. HCl, or a little over 0.2 per cent. It was found (Exps. 96, 99, 101, 102, 109, 112) that tutin solution kept at 37° C. in the presence of that amount of alkali completely lost its toxic power. In the first experiment the time allowed for the alkali to act on the tutin was one hour, and the three fish that were subjected to the action of the solution were unaffected after thirty hours. The fluid was filtered and three other fish introduced for twenty-seven hours, but without effect.
In other experiments the time allowed for the alkali to act upon the tutin was shortened successively to forty minutes, twenty minutes, ten minutes, and five minutes, but the result was the same in each case—the tutin had completely lost its toxic power.
In this connection the following experiment on a cat may be cited (Exp. 113): 8 c.c. of a 0.5-per-cent. solution tutin was treated with 1 gram of lime [Ca(OH2)] for one hour at 37° C., and thereafter for one day at room-temperature. The lime was removed by passing CO2 and filtering, and the fluid was evaporated to about 2 c.c., and injected under the skin of a cat weighing 2.4 kilograms. It displayed no symptoms, and remained well.
(e.) Influence of “Surviving” Organs on Tutin.
In considering the explanation of the relative immunity of birds to tutin poisoning by oral administration, it was suggested (vide ante) that the interposition of the liver in the usual path of absorption in mammals might account for some increase in the toxicity of the tutin in their case. To test the influence of the liver-substance on tutin, an experiment was carried out as follows: A rabbit was killed by chloroform, a canula was tied into the aorta, and the blood-vessels washed free of blood by a stream of warm saline. The liver and kidneys were rapidly excised, 9 grams of each weighed out, minced, and bruised rapidly in a warm mortar. To each mass 10 c.c. of a 0.5-per-cent. solution of tutin in normal saline was added. The mixtures, in small wide beakers, were placed in the oven at 37° C. for one hour. An equal volume of absolute alcohol was then added, to stop further action. The mixtures were then evaporated on a water bath, and kept there for some hours to coagulate the proteids. A watery extract of each was then made, filtered, and made up to 1,000 c.c. with tap-water. Three minnows were placed in each of these solutions, and the results were as follows:—
Liver and Tutin (Exp. 103): One fish showed symptoms at two hours and a third, and died in five hours and a half; one showed symptoms in two hours and a third, and died in six hours; and one showed symptoms in four hours and a half, and died in seven hours and three-quarters.
Kidney and Tutin (Exp. 104): One showed symptoms at an hour and a half, and died in four hours and a half; one showed symptoms at four hours and a half, and died in seven hours and three-quarters; and one showed symptoms in four hours and a half, and died in twenty-seven hours.
The course of events in the latter case is just similar to that of a 0.005-per-cent. solution of tutin, so that we may suppose that here the tissue had no effect on the tutin; but, on the other hand, it is difficult to avoid thinking that the liver-substance increased the toxicity of the tutin.
6. Action on Insects.
Insects are susceptible to the action of the poison. Weak solutions (0.1 per cent.) have no influence. A number of flies lived in perfect health
for a week on a solution of cane sugar in 0.1-per-cent. solution tutin (Exp. 115); but a 0.5-per-cent. solution quickly produced symptoms, and caused death in half an hour or so (Exp. 116).
The symptoms were very like those that characterize the action of the poison in higher animals—namely, an initial lethargy, followed by isolated tonic spasms of the wings and legs, exactly resembling the appearance met with in pigeons in the early stages of poisoning, when one or other wing is extended in rigid spasm for a moment; then marked clonic spasm of the wings recurring at intervals, and finally general convulsions, in which the insect tossed and buzzed about until exhausted. In the latest stages the flies lay on their backs, showing only occasional movements of the legs. After death, distention of the abdomen was an invariable finding, and in the case of a blowfly the exudation of a considerable drop of clear fluid from the proboscis occurred.
7. Action on Larvæ.
(Exp. 117.) The effect of the poison on maggots was tested by placing some mince that had become infected by blowflies in solutions of tutin of different strength for one hour, and then draining off the fluid. The experiment was controlled by a portion of the mince being placed in normal saline for the same period. On the day following, the maggots in the control had hatched out, and were very active; but in the mince subjected to the action of a 0.5-per-cent. solution tutin no movement was seen, and only one or two larvæ were found moving in the mince that had been soaked in tutin solutions of lesser strength.
Experiments that were made on full-grown maggots showed that confinement for forty-eight hours in tutin solutions of strengths graded up to 0.05 per cent. had apparently no influence. As immersion in a 0.05-per-cent. solution is about equivalent to a dose of 500 mlgm. per kilo, or 0.5 gram, tutin is seen to be very inactive.
8. Action on Molluscs.
(Exp. 114.) Two cockles of about the same size were placed one in sea-water and the other in a 0.5-per-cent. solution of tutin in sea-water, freshly prepared. Equal volumes of fluid were allowed. The cockle that had been placed in the tutin solution, in five minutes opened its shell and extruded its body. If touched it immediately withdrew, but came out again at once. It kept opening and closing its shell, but did not withdraw its body as the valves came together. It continued like this for twenty-four hours, responding more sluggishly to a touch as time went on, and then died. The control meanwhile had displayed no symptoms, but behaved in exactly the way the other had done when it, in turn, was placed in the tutin solution, and it also died.
9. Action on Infusoria and Amœbæ.
The action on infusoria and amœbæ was examined in the following way:—
A drop of hay-infusion, containing infusoria, amœbæ, monads, and bacteria, was placed on a slide; a drop of 0.1-per-cent. solution tutin was added, and the specimen observed for an hour. A drop of the infusion to which nothing had been added was used as a control. Another drop to which a drop of normal saline had been added was used to test the influence
of the saline in which the tutin was dissolved; and a fourth drop, to which a drop of a 0.1-per-cent. solution of quinine-sulphate had been added, was used for the sake of comparison (Exp. 118). In fifteen minutes no paramoecia were found moving in the quinine preparation, but monads were still in motion. In the three other specimens no change was observed. In thirty minutes all movement had ceased in the quinine preparation. In the tutin preparation paramoecia could still be seen moving, but their movements seemed to be somewhat irregular; amoebae, bacteria, and monads were still moving. In one hour one or two paramoocia could still be seen moving in the tutin preparation; they were by no means so numerous as at first, and rather difficult to find. Many of the monads were as active as ever, and amoebae were still throwing out pseudopods; but the preparation did not present the same appearance of general activity that it did at first. The saline preparations and the control were as active at the end of the experiment as they were at the beginning.
A further experiment (Exp. 119) was made by covering a drop of hay-infusion with a cover-glass, and then placing a drop of 0.1-per-cent. tutin solution on the edge of the cover. It was observed that such paramoecia as swam out into the drop of tutin solution remained in that drop, displayed irregular movement, and in a few minutes came to a standstill, and appeared to undergo internal disintegration, which reduced them to unrecognisable masses. The experiment was repeated with a drop of a 0.5-per-cent. solution tutin in distilled water, with similar results (Exp. 120).
It would appear, therefore, that paramoecia are injuriously affected by the poison, but not in so great a degree as they are by quinine. On amoebae, dilute solutions (about 0.05 per cent.) have very little influence. Monads are affected in a lesser degree than paramoecia.
10. Action on Bacteria.
(Exp. 121.) This was tested by placing pieces of fresh mince in test-tubes containing solutions of tutin of different strength (0.1 to 0.5 per cent.) and a piece in a test-tube containing normal saline alone. The tubes were left open to the air for twenty-four hours, and then corked and placed in an incubator at 37° C. Two days later the tubes were examined, and it was found that the tube containing saline was very offensive, but the tubes containing tutin solution were not at all offensive, and smelt rather like stomach-contents.
When subjected to microscopic examination, it was found that all the solutions contained bacteria, but in the slide prepared from the normal saline the bacteria were very much more numerous, and showed a greater variety of form and size. It would appear, therefore, that tutin has a deleterious action on certain forms of bacteria, but not on all.
That some forms of bacteria are certainly not affected by tutin is shown by the following observation: A 0.5-per-cent. solution of tutin in normal saline which had been allowed to stand for six months in the laboratory was examined microscopically, and found to contain fairly numerous large motile bacilli resembling B. subtilis. A green growth that had developed and formed a layer at the bottom of the bottle was found to consist of small rounded yeast-like cells tinged with chlorophyl.
11. Action on Yeast.
The action of tutin on the fermentation of glucose by yeast was studied in a few experiments (Exps. 122, 123, 124). It was found that fermentation
proceeded vigorously in the presence of even 0.4 per cent. tutin. Some slight differences in the rate of fermentation that were noted were probably due to differences in the amount of yeast added. (For details of experiments, see protocols, Exps. 122, 123, 124.)
12. Germination of Seeds.
The action of tutin on the germination of seeds was tested by soaking mustard-seeds in solutions (in normal saline) of various strengths (0.1, 0.2, 0.3, 0.4, and 0.5 per cent.) for twenty-four hours (Exp. 125). The seeds were then sown on pieces of felt in separate tin boxes, appropriately labelled, and kept moist with water. An equal number of seeds was sown on each piece of felt, and the experiment was controlled by seeds that had been soaked for the same length of time in normal saline. The day following, all the seeds had germinated, and they were therefore replaced in their respective tutin solutions for a further period of twenty-four hours, and then sown again. During the next week the progress made by the seedlings was observed, and the effect of the tutin on their development noted.
It was found that the seeds progressed more or less in order of the weakness of the solutions in which they had been soaked: thus, the growth of those that had been soaked in normal saline, in 0.1 per cent., and in 0.2 per cent. tutin was the most vigorous, while those that had been soaked in 0.3, 0.4, and 0.5 per cent made slow progress, and all the seedlings in these lots did not continue to grow. In a fortnight, however, little difference was to be noted in any of the seedlings that had developed at all, one or two of the 0.5-per-cent. seedlings that were growing being as well developed as those that had been soaked in saline.
This result may possibly be due to the effect previously noted on bacteria certain forms of bacteria being necessary to the growth of plants. From experiments on invertebrates, bacteria, &c., it may be inferred that tutin is toxic to any given form of life in proportion to its complexity of organization. Wherever a nervous system is sufficiently developed, the toxic action is greatest; but that it has some injurious influence on primitive protoplasm may also be seen from its effects on paramoecia, bacteria, &c.
13. Action of Tutin on Tissues.
From the symptoms observed when studying the general action of the poison on mammals, it seemed very evident that the nervous system was the part chiefly affected. Nervous tissue will be considered later, and the action of tutin on other tissues will be recorded first.
(a.) Action on Primitive Protoplasm.
As is shown in Exp. 118, tutin, as compared with quinine, is not a strong protoplasmic poison. That it has some deleterious action on living protoplasm is shown by the experiments on putrefactive bacteria, on the germination of seeds, and on paramoecia (Exps. 121, 125, 119), though it is doubtful if the action here is more than might be exerted by any foreign substance.
(b.) Ciliated Epithelium.
The action on cilia was tested in the following way: A few bars from the gill of a cockle were placed on a slide in a drop of sea-water, and examined with a power of 80 diameters. In the first experiment, the preparation was covered with a cover-glass, but in the later experiments coverglasses
were not used, in order to allow of ready interchange of O2 and CO2. The results are shown in Table VIII (Exps. 126 to 134 inclusive).
|Exp. No.||Source of Material.||Strength of Tutin.||Effect.||Time.||Remarks.|
|126||Cockle-gill Same preparation||0.1 in saline||No change||7 min.||Control specimens in normal saline continued active.|
|"||0.2 "||Retarded (?)||4 "|
|0.3 "||Ceased||8 "|
|127||Cockle-gill||0.3 "||Slower 1 " Ceased||1 "|
|128||"||0.2 "||Practically ceased||7 "||Movements restored by 1 in 1,000 KOH.|
|129||"||0.2 "||No change||9 "|
|Same preparation||0.3"||Less active||10 "|
|"||0.5 "||Still moving||15 "|
|130||Cockle-gill||0.2 "||Slowing||7 "||Accelerated by infusion with sea-water.|
|131||Rock-oyster gill||0.5 "||Slight retardation (?)||5 "|
|Still active||1 ½ hours.|
|132||Cockle-gill||0.5 "||Exaggerated (?)||2 min.|
|Still active||1 ⅓ hours.|
|133||Ciliated epithelium of frog's gullet||0.1 "||No change||1 hour.|
(Exp. 134.) The rate of progress of a small fragment of cork along the surface of the gullet was noted before and after applying tutin (0.3 per cent. in saline). Average rate before = 50.4″ for 1 cm. travelled; average rate after = 34.8″ for 1 cm. travelled. The method presented many difficulties, as it is impossible to keep the fragment moving on the same line each time, and the mucus secreted clogs its movements.
From these results it is impossible to draw a definite conclusion, for although in some specimens taken from sea-water animals the ciliary action ceased on the application of tutin, in other specimens from the frog's gullet tutin had little or no action.
(c.) Action of Tutin on Striped Muscle and Nerve-terminations.
This was tested by determining the strength of stimulus just sufficient to cause contraction of a nerve-muscle preparation before and after the application of tutin. The preparation (gastrocnemius - sciatic) was made from the frog. An induction-coil with one Daniell cell and mercury-key was used. The muscle in each case was placed first in normal saline (0.75 per cent.) and the strength of the minimal stimulus determined. The tutin solution was then substituted for the saline, and the excitability tested from time to time.
Four experiments were made (Exps. 135 to 138 inclusive). In three of these the one muscle was placed in 0.025, 0.05, and in 0.5 per cent. tutin solution, while the other muscle was placed in normal saline. In another experiment the muscle of one preparation was placed in 0.5-per-cent. solution tutin, and the nerve of the other in 0.5-per-cent. tutin. (Exp. 136).
In the first experiment (Exp. 135)—0.025 per cent. tutin—there was a diminution in excitability; thus, the muscle in saline contracted with the coil at 18cm., three hours after the experiment began, while the corresponding muscle in tutin contracted only at 15 cm.
Where 0.05-per-cent. solution tutin was used (Exp. 137) the muscles were equally excitable after two hours, but the muscle in the tutin had been the more excitable of the two before immersion in the tutin. Thus, before tutin was applied it contracted at 30.5 cm., and two hours after the application of tutin at 25.5 cm., while the control contracted at 25.5 cm. and at 25 cm.
Another experiment where the muscle was placed in 0.05-per-cent. solution tutin gave the following results (Exp. 138): Before immersion in tutin solution—A contracted at 36 cm.; B (control) contracted at 32 cm. Two hours after immersion in tutin solution—A contracted at 21; B contracted at 23.
In the experiment with 0.5-per-cent. solution tutin (Exp. 136) the minimal stimulus before the application of tutin for A was 28 cm., for B 34 cm. The muscle of B (control) and the nerve of A were then placed in the tutin solution, and an hour and a quarter afterwards A contracted at 33 cm. and B contracted at 27.5 cm.
(For details of these experiments, see protocols.)
The inference to be drawn from these few experiments is that tutin has no striking action on nerve or muscle fibre; but there seems to be a diminution in the excitability in the muscle placed in tutin solution, due either to an action on the nerve-terminations or to an action on the muscle fibre. This would help to explain the weakness and loss of tone seen in the later stages of tutin poisoning, especially well seen in frogs.
It is possible that the action is one on the nerve-endings, for instances are known of a poison (e.g., strychnine) acting as an irritant to nerve-cells, and having at the same time a paralysing action on motor-nerve endings. Nussbaum (27) states that picrotoxin in small doses stimulates, and with slightly higher doses depresses, the activity of the peripheral nerve-terminations. The same may be the case with tutin.
14. Action on the Different Systems.
(a.) Alimentary System.
Salivation is a marked feature in animals poisoned by tutin. It is an early symptom, and usually precedes the vomiting, though it may be present when vomiting is absent, as in Exps. 1 and 2. It is most profuse where the dose is large, and in these cases is a source of danger in the later stages by obstructing the respiratory passages. As a rule, it diminishes when vomiting has occurred, and is not a marked feature in those animals (rabbits and guinea-pigs) that do not vomit. Probably, therefore, it may be an exaggeration of the salivation which usually precedes the act of vomiting. The saliva is thick and ropy, and hangs from the mouth in long tenacious strings. Saliva of this consistence suggests stimulation of the sympathetic.
Vomiting is also an early symptom. It is not an invariable symptom, being present in only two of the cats and in three or four of the pigeons experimented upon. It is more likely to occur if the dose be small.
The salivation and the vomiting could both be explained as due to stimulation of the vomiting-centre in the pons. That it is due to stimulation of the centre is shown by the fact that it occurs when the poison is
given hypodermically. But in ordinary cases of poisoning by the tutu plant there may be irritation of the gastric mucosa as well. Christie's assertion (11) that tutin is excreted by the stomach, and so causes vomiting by local irritation, is unsupported. No tutin could be extracted from the gastric and small-intestine contents of a rabbit which had been killed by intravenous injections of large doses of tutin during a blood-pressure experiment.
Stomach.—In cases of herbivora poisoned by tutu, accumulation of gas in the stomach is a marked feature. This is probably due to the rapid fermentation of the leaves ingested, though possibly it may be due, in part, to air swallowed during the convulsions.
To test the fermentability of tutu-leaves, a mash of minced leaves with a little water was placed in an incubator at 40° C., and examined after the lapse of an hour. On stirring the mixture it was seen to be permeated with bubbles of gas, but it was not determined how far this was due to the expansion of imprisoned air.
In the experiments with the pure substance, distension by gas was not observed. Pure tutin and artificial gastric juice when incubated in a fermentation-tube gave no appearance of gas-development.
The action of hydrochloric acid in hydrolysing tutin has already been referred to under “Fishes.” It was found that the toxicity was slightly increased.
Intestines.—The action on peristaltic movement was not investigated experimentally, but from observations made on animals killed by tutin, where the abdomen was opened immediately, it was clear that there was no diminution of peristaltic movement. That it was even increased was inferred from the frequency with which defaecation occurred. The stools were often loose, and were expelled with some violence.
(b.) Haemopoietic System.
In one experiment, where a rabbit received a fatal dose of tutin and died in one hour, blood-films were taken before the injection of tutin and just before death (Exp. 139). The only point of difference seemed to be that the leucocytes appeared fewer in number after tutin. They seemed to be quite normal in regard to granules and staining-power.
(c.) Circulatory and Respiratory Systems.
Previous observers have noticed that the tutu poison has no depressing influence upon the circulation. Christie (11) found that the heart continued beating after the respiration had ceased. In one of his experiments (on a cat) he opened the thorax immediately on the cessation of all movement, and observed the heart to beat for twenty-two minutes after the last respiratory gasp. In his own case, where he took 9 grains of an extract made from the leaves, the pulse-rate rose to 102, and this observation led him to conclude that the action of the poison was to accelerate the beat of the heart. In attempting to bleed poisoned animals by opening veins and by slitting the ears he found that the blood did not run readily, and concluded that the arterioles were contracted. He noticed, also, what had been observed previously in cases of accidental poisoning—that at first the respirations were increased in frequency and in force, and that later they became feeble and irregular, and finally ceased before the heart stopped beating.
Marshall (24) found that tutin diminished the number of heart-beats and increased the frequency of respiration.
As coriamyrtin belongs, pharmacologically, to the picrotoxin group, and is therefore closely allied to some of the decomposition-products of digitalin—e.g., digitaliresin (30)—it was expected that tutin, which is so closely related to coriamyrtin, might show the tonic action on the heart that is characteristic of the digitalis bodies. In frogs and in mammals, where the heart was examined after the breathing had ceased, it was found invariably to be still beating. In one experiment on a cat (Exp. 153) the heart-beat was distinctly audible with the stethoscope for two minutes and a half after respiration had ceased.
As it had been observed that tutin has apparently no action on striped muscle (Exps. 135 to 138), and as no proof was forthcoming that it has any action on unstriped muscle, it was not considered likely that it would manifest any direct action upon cardiac muscle. As it might, however, have some action on the extrinsic and intrinsic nervous mechanism of the heart, it seemed best to begin the study of its action upon the circulation by observing its effect upon the heart of a pithed frog, where one has to consider muscle and intrinsic ganglia only. A number of experiments were undertaken to determine this. Tracings * of the frog's heart-beat were taken in the usual way with a lever arranged to write against a blackened revolving cylinder. In some cases the tutin solution was painted on the surface of the heart; in others it was injected into a vein or into the heart itself. The results obtained show that the rate of the beat was slightly diminished: thus, in Exp. 140 the rate fell from 37 to 35 beats per minute; in Exp. 141 from 32 to 30.8, then to 28.8, and then to 26.4 per minute; and in other experiments from 20 to 14 and from 20 to 16.
After injection of tutin the force of the heart-beat was increased, and the beat was not impaired even with a concentration of tutin solution greater than could possibly be reached by absorption of the poison from the stomach in cases where the plant has been eaten.
An irregularly beating heart was generally improved, the beat becoming regular. So that it may be concluded that tutin has a purely beneficent influence on the heart's contraction.
Action on Mammalian Circulation.—This was studied by taking tracings of the blood-pressure from one or other carotid artery, and noting the effect of injections of tutin solution into the jugular vein. The animal was anaesthetised (chloroform and paraldehyde) during the experiment, and was not allowed to regain consciousness. It was found that injections of tutin solution (0.5 per cent.) caused a rise of pressure: thus in Exp. 143 the initial pressure was 3 in.; in the course of an hour, during which the animal received nearly 10 mlgm. tutin, the pressure rose gradually to 4 ½ in. In some cases the vagi were divided before the drug was injected, but without affecting the result. The rate with which the blood-pressure rises is proportional to the size of the dose given. In one experiment (Exp. 145), where the abdomen had been opened and its contents handled, a reflex cardiac inhibition seemed to be set up, for the tutin injection in this case caused a marked fall in pressure, which was followed by a rise on division of the vagi. It appears, therefore, that tutin increases the
[Footnote] * As the tracings had to be sent to Edinburgh with the original thesis, it is impossible to reproduce them here, and this portion of the work has therefore been summarised, and technical details have been omitted.
excitability of the medullary centres, and the stronger reflex effects from these usually lead to a rise of blood-pressure; but in cases where some reflex cardiac inhibition already obtains, this is increased by tutin, and a fall in pressure results. That tutin acts specially on the centres in the medulla was also shown in several experiments where Traube-Hering curves and Cheyne-Stokes type of respiration occurred simultaneously. When twitching and general convulsions set in, the blood-pressure curve became markedly irregular, due probably to mechanical obstruction to the flow through the vessels.
The action of tutin on the vagus-endings in the heart was investigated. The strength of stimulus just necessary to inhibit the heart slightly and cause a fall of blood-pressure was determined both before and after the injection of tutin. No marked difference was obtained, so it was concluded that tutin has no influence on the excitability of the nerve-ending in the heart.
In some of the later experiments a spring manometer was used to measure the blood-pressure instead of the ordinary mercurial manometer. This, by recording each heart-beat on the tracing, enabled the rate to be accurately determined. It was found that there was no increase, but in rabbits the rate is already so high that one could hardly expect much change.
A curious periodicity in the heart's action was observed in some cases towards the end of the experiment when the animal was dying, one or two beats being missed at each inspiration. The explanation of this was not at first clear, but in a subsequent experiment on a cat (Exp. 153), where the heart was auscultated as the animal was dying and breathing in the way shown in the tracing, it was noticed that towards the end of each inspiration the heart missed two or three beats. The explanation of this seemed to be that as the respirations were infrequent and deep, and the respiratory passages obstructed by quantities of mucus and saliva, the deep forcible inspiration created so much negative pressure in the thorax that the heart was unable to beat until the pressure was relieved by expiration. That the variation was due to respiratory influence, and not to any action of the drug upon the circulation, was shown in the case of this cat by the fact that when respiration had ceased the heart continued to beat regularly for two minutes and a quarter.
With regard to Christie's statement that the arterioles are contracted, no direct experiments were made, but some perfusion experiments on frogs showed that there was no marked or constant change. Further, the action on the heart is sufficient to account for the rise in blood-pressure. Were the arterioles also constricted, one would expect a much higher rise than occurs. Any influence that tutin does exert on the calibre of vessels is exerted through the vaso-motor centre in the medulla, as seen in the Traube-Hering curve already mentioned.
Action on the Respiratory System.— The rate and amplitude of the respiratory movement is markedly increased by tutin. After the first injection in the anaesthetised animal the rate may be doubled. During convulsive seizures the spasm of the respiratory muscles is very marked, and respiration appears to be brought to a complete standstill—e.g., in a cat (Exp. 3), during an interval of four minutes and a half, no visible or audible sign of breathing could be noticed. Doubtless this is a frequent cause of death in animals poisoned by a large absorption of tutin.
That respiration ceases before the heart was a constant observation. It was seen in fishes and in all animals where such an observation could be made.
Breathing of the Cheyne-Stokes type occurred in some experiments, the climax of the period of respiratory activity corresponding to the highest point of the Traube-Hering curve, which appears simultaneously.
(d.) Urinary System.
Micturition was a marked feature in cats poisoned by tutin. It occurred voluntarily several times before convulsions occurred, and during the convulsions (and this is true also of rabbits and guinea-pigs) involuntary discharges of urine occurred. In the majority of cases the bladder was found distended after death, even where a free discharge of urine had taken place during the convulsions. This points to an increased secretion of urine. Attempts were made to estimate the rate of flow in anaesthetised animals (Exps. 144, 145), but without satisfactory results.
This increased secretion pointed to the possibility of tutin being eliminated in the urine, and so an attempt was made to recover tutin from the urine of poisoned animals. The urine was collected by incising the bladder after death. It was evaporated to dryness and exhausted with ether, but no crystals of tutin could be detected on evaporating off the ether. The residue from the ethereal solution was in one case dissolved in saline and injected under the skin of a frog (Exp. 151), but with negative results. The urine was in several cases examined for abnormal constituents, but none was found.
(e.) Genital System.
Many of the animals experimented upon were pregnant females in different stages of pregnancy, but in no case was abortion observed, although the animal was under the action of the poison long enough for it to occur.
(f.) Nervous System.
The symptoms that appear in animals poisoned by tutin point clearly to the central nervous system as the part of the body that is specially affected by the action of the poison. The two cardinal symptoms are convulsions and a dulling or blunting of the mental faculties that in the early stage makes the animal appear dazed and stupid, and in the later stages passes into actual coma.
Convulsions.—The convulsions are obviously of central origin. They bear a close resemblance to an ordinary epileptic fit, which is generally held to originate in the cortex. Whether the tutin convulsions originate in the cortex or not, it is certain that the nerve-cells in the basal ganglia, pons, medulla, and cord are profoundly affected. This is shown by the action on the vomiting centre, respiratory centre, and cells of the cord. Among the first symptoms to appear are twitching of the ears, blinking of the eyes, and movements of the lips. These are followed by jerking of the head and movements of the fore and hind limbs. These movements may be attributed to irritation of the cells of either the upper or the lower motor neurones. In order to determine more accurately the site of action the following experiment was made:—
(Exp. 152.) The right cerebral hemisphere was exposed in a cat and a large dose of tutin injected hypodermically. When general convulsive movements had developed, the right cerebral hemisphere was removed. Now, had the convulsions been cortical in origin, one would expect them to have ceased on the left side of the body; but they continued as before,
affecting both sides equally. The left hemisphere was then removed, but the movements were unaffected, and continued on both sides equally. So it may be concluded that the convulsions may originate in the cells of the lower neurones, or, at any rate, that the upper neurone is not necessary for their development. Further observations made in this experiment lend support to this suggestion. After the cerebrum had been removed, the cord was divided at the level of the fifth dorsal vertebra. Four minutes afterwards tonic and clonic spasms of the hind limbs were observed, and continued intermittently for a quarter of an hour. As the cells of the cord were now completely cut off from the higher centres, these movements must have originated in the neurones of the spinal cord itself.
Post Mortem.—The chest was opened just after all respiratory movement had ceased, and the heart was seen to be still beating. The bladder was full. The cerebral hemispheres were found to be entirely removed. The cord at the seat of section was examined, and it was found that division was complete, but there had been some crushing of the tissue on either side of the section, and the dura mater was still intact. Although satisfied that division was complete, it was thought that the crushing of the cord may have caused irritation, which might possibly be held to account for the movements observed in the hinder part of the body. It was resolved, therefore, to repeat the experiment. This was done as follows:—
(Exp. 153.) A cat was chloroformed, the cord exposed, and completely and cleanly divided in the mid-dorsal region. Five mlgm. of tutin were then injected into the peritoneum, and chloroform anaesthesia discontinued. Ten minutes later no reflex could be elicited from the left hind foot, while the left fore foot responded normally. Fifteen minutes after the injection of tutin twitching of the ears and jerking of the head began, and became very distinct at twenty-four minutes. Two minutes later a convulsive movement of the fore part of the body occurred, and was accompanied by a movement of the tail. These convulsive movements of the fore part of the body continued for nearly an hour, and the animal died an hour and seventeen minutes after the injection of tutin. About half an hour after the cord had been divided and tutin injected, reflexes could be obtained from the hind limbs; and ten minutes later a stimulus applied to one hind limb caused movements of both. During the last half-hour of life the movements of the tail became more and more marked. They generally appeared just at the beginning of each convulsive seizure of the fore part of the body, but they also occurred independently. Sometimes the whole tail was moved from the root, at other times there was only a slight movement of the tip. One hour after section of the cord convulsive movements (tonic and clonic spasms) of the hind limbs occurred, and were accompanied by defaecation and erection of the hairs of the tail. These convulsive movements were frequently repeated. By this time the fore part of the body had become almost quiescent, but the spasms of the hind limbs increased in severity until death. It was in this experiment that the heart was observed to beat intermittently during the long-drawn, obstructed inspirations. When respiration had ceased, it continued to beat regularly for two minutes and a quarter.
The conclusion to be drawn from these two experiments is that the convulsions that occur in tutin poisoning may originate in the lower centres of the pons, medulla, and cord. The action on the cord is a late appearance, and this corresponds to the finding of Gottlieb (28) that picrotoxin can elicit convulsions below a section of the spinal cord, and that these appear later
in the hinder part of the body. This result does not agree with the statement made by Marshall (24) that the spasms in tutin poisoning are not produced below a section of the cord.
Like other convulsants, tutin raises the excitability of the reflex arc. This is shown in the last experiment, where crossed reflexes could be obtained soon after section of the cord. This rise in excitability might explain the severity and frequent repetition of convulsive seizures when once they begin, for it is known that in strychnine poisoning, where the reflex excitability is raised, the afferent, probably painful, impulses coming from the convulsed muscles originate further convulsions, and thus a sort of vicious cycle is set up. In this connection it may be noted that it has long been known to farmers that if “tooted” sheep are left undisturbed no symptoms may appear; but once the symptoms are initiated—say, by the bark of a dog—they continue, and are from the outset severe. The explanation given above would account for the apparently sudden onset of severe symptoms.
It seemed of interest to determine whether the reflex time was shortened. Florence Buchanan (29) and others found that strychnine in small doses did not perceptibly shorten the reflex time. It was impossible in this case to use the same elaborate method, so recourse was had to the simple but less accurate method of Turck. The results of the experiments, with details of the method, are given in the protocols, but they may be shortly summarised here.
Six experiments were made. In three of these (Exps. 154, 156, 157) the reflex time was shortened from 6 to 4, from 6 to 5, and from 6-1 to 3 respectively; while in three others (Exps. 155, 158, 159) the time was lengthened from 8 to 12.2, from 8-3 to 22, and from 8-5 to 44. From these results it will be seen that no definite conclusion could be drawn. The one thing that did appear in these experiments was that the amount of movement was increased after the injection of tutin, the stimulus as a rule now causing general movements of the body. Thus, though the effect of tutin upon the reflex time remained doubtful, its effect in increasing reflex excitability seemed to be clear.
In frog 157 the injection of tutin caused a typical convulsion, and in this case it was found that the optic lobes had not been pithed. In cases where the pithing destroyed all parts but the cord no convulsions occurred. Although this differs from what was found in mammals where tutin produced spasms below a section of the cord, it agrees with Gottlieb's(28) finding that in picrotoxin poisoning in frogs convulsions did not occur below a section of the cord.
The second important symptom of tutin poisoning is the comatose condition that is so invariable an accompaniment. This occurs in various degrees according to the dose, and it deepens as the case advances. In the recorded cases of poisoning of human beings, referred to above, complete loss of consciousness and subsequent loss of memory was frequently observed. Loss of memory appears to occur also in lower animals, for shepherds have observed that a “tooted” lamb which has recovered does not know its own mother, and the ewe may be seen following her lamb and striving to excite recognition.
Another proof that abrogation of sense-perception obtains is the marked absence of any evidence of pain. The animals utter no cry of pain, although the convulsions are very severe, and would, were this condition not present, be most painful.
From the examination of sections of the cerebrum, medulla, and cord there is evidence of great congestion of these parts in tutin poisoning. The central nervous system was examined in several cases after death, and congestion of the membranes was found in all. In one case the grey matter of the cord when seen on section was distinctly reddish. Pieces of cord, medulla, and cerebrum were fixed in 8 per cent, formol, and sections were made and stained by Nissl's method, and also by Muir's eosin and methyline-blue method. On microscopic examination the Nissl granules showed no obvious change, but there was very evident congestion, shown especially well by Muir's method. The capillaries and small vessels seemed more numerous than normal, because they were rendered visible by being crowded with red blood-corpuscles. At several places there were collections of corpuscles, pointing to small extravasations of blood. These changes were present in the grey matter of the cord, medulla, and cerebral cortex. The appearance at once suggested that more or less permanent damage would have resulted had the animal lived. It has been observed in cases of tutu poisoning that sometimes the victim does not completely recover, a permanent mental alienation remaining as an after-effect.
Lauder Lindsay (3), in one case of poisoning he records, in which convulsions were a prominent symptom, notes that the subject never completely recovered, “there remaining to this day a peculiar form of nervous irritability not observable prior to this toot-poisoning.” The present writer has knowledge of another case, although, unfortunately, no authentic details are available, where two children were poisoned by tutu. They were both very seriously ill, and one died. The other recovered, but incompletely, mental enfeeblement and a squint remaining as after-effects. Sequelæ of these kinds may possibly be explained as resulting from permanent damage done to the nervous tissue by the extreme congestion and the small hæmorrhages which occur in the cerebral cortex as well as through-out the whole grey matter of the central nervous system.
Sympathetic Nervous System.—This was not specially examined by experimental methods, but that the nerve-cells here were also affected was shown by the viscid character of the saliva secreted by cats, by the dilatation of the pupil that occurred during convulsions, and by the erection of the hairs of the tail.
In short, it is probable that tutin affects every kind of nerve-cell; the fact that the medulla is seen to be specially affected being due to the sensitiveness of the cells in that region.
Effect on the Pupil and Conjunctiva.—When dropped into the eye of a rabbit, tutin solutions cause no local irritation and no change in the size of the pupil (Exps. 160, 161). The same holds good for the cat (Exp. 162) and for the excised eye of the frog (Exp. 163). During convulsions in the cat and in pigeons dilatation is well marked.
The action of coriamyrtin was studied in the same way, because it is said by Riban (15) to cause contraction of the pupil in rabbits when applied locally. It was found to cause contraction in the excised eye-ball of the frog, but the results of applying it to the eye of a rabbit and of a cat were negative (Exps. 164, 165).
(g.) Action of Tutin on General Nutrition.
It is not an uncommon opinion among farmers that animals that eat the plant in moderation thrive well on land where tutu abounds. To test whether tutin had any injurious influence on general nutrition, a young
rabbit (Exp. 166) was given small doses of tutin by the mouth every third or fourth day for nearly two months, the dose being gradually increased up to 7 mlgm. per kilo (6 mlgm. per kilo is a fatal dose by the mouth in rabbits). During this time the animal grew and developed normally, and it increased in weight from 777 grams to 1,154 grams. This shows that tutin does not retard growth.
Another experiment was made on a guinea-pig (Exp. 167), the animal being treated like the rabbit. A second guinea-pig, as a control, was kept in the same hutch, and abundance of food was, supplied. In thirteen days the animal subjected to the influence of tutin gained 16 grams, and the control in the same time gained 38 grams. The animals, however, were not of the same age; the control, being younger, apparently gained more.
From these data it looks as if tutin had no injurious influence on general metabolism (cf. Exp. 15, on a pigeon).
15. Fate Of Tutin In The Body.
It is impossible to trace in the body the fate of a substance which, like tutin, contains only C, H. and O; but that the tutin is not rapidly destroyed or eliminated was shown by the following experiment (Exp. 13): A guinea-pig was given by mouth on alternate days a dose of tutin which was near the minimum lethal dose; thus, 1-5 mlgm. per kilo was given on the 17th, on the 19th, and on the 22nd February. No symptoms appeared, so the animal was able either to oxidize or to excrete this amount (1.5 mlgm. per kilo) in two days. On the 24th it received 2 mlgm. per kilo, and a similar dose was given on the 26th. On the 27th it was found dead. From this it appears that a guinea-pig is unable to dispose of 2 mlgm. per kilo within two days. Enough tutin must have been still present in the body on the afternoon of the 26th, when the second dose of 2 mlgm. per kilo was given, to raise the total amount present to the lethal dose.
16. Immunity Or Tolerance.
Several experiments were made with the object of determining whether tolerance of the effects of the poison can be established.
A pigeon (Exp. 15) that had been subjected to gradually increasing doses by mouth over a period of ten days withstood 16 mlgm. per kilo. The minimum lethal dose in pigeons by oral administration is about 10.25 mlgm. per kilo. A second pigeon (Exp. 17), treated in the same way for over three weeks, succumbed to 12 mlgm. per kilo. A guinea-pig (Exp. 167), which was being treated in the same way, and had received several small doses, was killed by a dose of 7.5 mlgm. per kilo given by mistake, so that no great degree of tolerance had been established. A rabbit, already referred to above when considering the effects on general metabolism, received small doses, gradually increasing to 7 mlgm. per kilo, over a period of two months. A dose of 8 mlgm. per kilo then caused severe symptoms; so that it would appear that a slight degree of tolerance had developed, for the lethal dose, by oral administration, in rabbits is about 7 mlgm. per kilo. Five days later this rabbit succumbed to a dose of 11.6 mlgm. per kilo per os.
The general conclusion reached was that tolerance can be established only in a very slight degree, if at all.
Schmiedeberg (30) placed coriamyrtin in the picrotoxin group of nerve and muscle poisons. This group is closely allied to the camphor and digitalin groups, which include a number of the strongest non-nitrogenous poisons found in the vegetable kingdom. Other members of the group are circutoxin, a resinous substance derived from the water-hemlock; œnanthotoxin, from the water-dropwort; digitaliresin and toxiresin, decomposition products of digitalin and digitalein respectively; and oleandresin, from oleandrin. These substances, by their action on the medulla oblongata, all cause general and respiratory convulsions, slowing of the pulse, and a rise in blood-pressure.
Tutin, as has been shown, possesses these actions common to the group. Some experiments were made to determine its toxic power as compared with picrotoxin and coriamyrtin (a very small quantity of the latter—11 mlgm.—dissolved in absolute alcohol was kindly supplied by Mr. Aston, of the Agricultural Department of the New Zealand Government; he, in turn, had obtained it from its discoverer, Monsieur Riban).
Tutin compared with Picrotoxin.—In the experiments on fishes already cited it was found that picrotoxin was more lethal than tutin when equal percentages of the two were used, and in equimolecular solutions it proved much more lethal. In mammals, on the other hand, tutin was found to be more fatal than picrotoxin. In Exp. 4 a cat received 0.375 mlgm. per kilo of tutin; it developed severe symptoms, but recovered. Two days later it received exactly the same weight of picrotoxin (Exp. 168) without the development of any symptoms. The solution of picrotoxin used had been made seven months previously, and, lest it had deteriorated, a fresh solution was made and the same dose repeated five days later (Ex. 169); beyond defæcation and occasional swallowing-movements, no symptoms appeared. It was therefore concluded that tutin was more poisonous to mammals than picrotoxin. Following the procedure adopted in the case of fishes, the two were then compared in equimolecular strengths, and 0.375 × 883/336 mlgm. per kilo given hypodermically to the same cat (Exp. 170).
It developed twitching, vomiting, and other symptoms resembling those seen with tutin. Only one general convulsion occurred, however, whereas the dose of tutin had caused very numerous convulsions. It recovered in a shorter time, and the whole effect was much less severe; so that, in contradistinction to fishes, mammals may be said to be more susceptible to poisoning by tutin than by picrotoxin.
Tutin and Coriamyrtin.—The quantity of coriamyrtin mentioned above was insufficient for an extended series of experiments, and only one cat was subjected to its influence (Exp. 171). The symptoms were very similar to those of tutin poisoning; twitching and convulsions were very marked. In the convulsions the tonic spasms were not so prolonged as in the case of tutin, the clonic element being more in evidence; emprosthotonos appeared instead of the opisthotonos, which is so invariable a feature of tutin convulsions. Salivation, vomiting, respiration, erection of hairs of tail, and dilatation of the pupil were all present, as in tutin poisoning. A strict comparison of its toxic power can hardly be drawn from one case; but if the time taken to kill be proportional to the toxic power, tutin is the more lethal of the two, for 3 mlgm. per kilo of tutin killed a cat (Exp. 1) in thirty-one minutes, while this cat with the same dose of coriamyrtin died in forty-two minutes.
(Exp. 172.) Twelve mlgm. per kilo proved fatal to a frog. It displayed the symptoms seen in tutin poisoning. The time of death could not be noted, so no strict comparison can be made with tutin. The minimum lethal dose of tutin in frogs is between 10 and 11 mlgm.
On the whole, the impression left on one's mind is that there is comparatively little difference in the toxic power of the two substances; but the mental effects seemed more marked in the case of the cat poisoned by tutin.
18. Action Of Remedies.
Although not strictly within the scope of the title, of this paper, notes of some experiments made to show the influence of remedial measures will be included here. These experiments were made from time to time while the physiological action of tutin was being examined, and before the experiments on the action of alkalis on tutin had been undertaken. They are necessarily, therefore, incomplete; but some points have been investigated and some observations made which it is hoped may prove of value in the treatment of cases of poisoning.
In tutu poisoning various remedies have been suggested from time to time, and the rationale of some of these is difficult to understand. With shepherds, bleeding is a favourite method of treatment. It is usually done by slashing the ears or tail, or by incising the roof of the mouth. It is said to be of special advantage in young sheep, but in older sheep it is regarded by some as being dangerous, and as tending rather to hasten the end than to promote recovery. Carbonate of ammonia is also used, a lump about the size of a walnut being dissolved in water and poured down the animal's throat.
In 1870 Hughes advocated the use of lime as an antidote. He was led to do this from the observation he had made that lime destroyed the activity of the poison. Cases in human beings have been treated with lime, and, it as said, successfully.
The Maoris depended largely upon partial asphyxiation as a means of treatment. This was effected either by holding the patient under water till he was nearly drowned, and repeating the immersion as soon as he showed signs of returning life, or by suspending him head downwards over the smoke of a fire. Another method (31) was to bury the patient in the ground up to the neck, apparently with the object of restraining the convulsive movements.
Professor Marshall (32), in a report made to the Agricultural Department of the New Zealand Government, recommends bleeding and the intravenous injection of chloral-hydrate. In connection with the use of chloral-hydrate it may be noted that Crichton Brown (34) states that he was able, by the administration of chloral-hydrate, to prevent death in a rabbit which had received five times the minimum lethal dose of picrotoxin.
It will be noticed that in a rabbit (Exp. 174) which received a lethal dose (3 mlgm. per kilo hypodermically) death was prevented by 0.6 gram of chloral-hydrate. It should be stated that the chloral was given first by the rectum, and the tutin administered hypodermically as soon as the anæsthetic effect of the chloral was established, so that every chance was given to the action of the chloral. In two rabbits where 4 mlgm. of tutin per kilo was given (Exps. 173, 175) death occurred. The one received 1 gram of chloral per rectum in one dose, and the tutin hypodermically immediately afterwards; it died in six hours, in tutin convulsions. The
other received 0.6 gram chloral per rectum, and while asleep, ten minutes later, the tutin was given hypodermically; symptoms of tutin poisoning came on in less than two hours, and continued for an hour and a half; as the animal seemed likely to succumb, a further dose of 0.6 gram chloral was given, and in about a quarter of an hour the symptoms subsided, and the animal apparently recovered; next day it was suffering from diarrhœa, and the following day was found dead; possibly it died more from the after-effects of the chloral than from tutin, and it looked as if a little careful nursing would have led to its recovery.
The results of some experiments made with chloral as a remedy in tutin poisoning are given in Table IX.
|Exp. No.||Animal.||Dose of Tutin in Mlgm. per Kilo.||Usual Result.||Remedy used.||Result.||Remarks|
|167||Guinea-pig||7.5||Death within 1 hour||Chloral. 0.5 grm||Died in 45 hours||Died apparently from chloral.|
|173||Rabbit||4||Death within 2 ½ hours||0.6 grm., followed by a second dose of 0.6 grm.||Died between 24 and 48 hours|
|174||"||3||Death within 3 hours||0.6 grm||Recovered completely|
|175||"||4||Death within 2 ½ hours||1 grm given simultaneously||Died in 6 hours||Tutin symptoms.|
A guinea-pig (Exp. 167) which had received 7.5 mlgm. tutin per kilo by mistake, and was then given chloral per rectum, died in a somewhat similar manner. It recovered fully from the tutin convulsions, but became paralysed in its hind quarters, and died on the second day after the tutin was given. It also had diarrhœa.
Frogs (Exps. 177, 178, 179) which were given just over the lethal dose of tutin (11 and 12 mlgm. per kilo) were not saved by chloral in moderate doses given hypodermically at the same time. Control Exp. 176.
On the whole, the use of chloral seems to be justified, and in cases of poisoning in man, where the symptoms can be more accurately observed and interpreted, and dangers more easily circumvented, it should prove of great value. The beneficent action of tutin on the heart would allow of the use of larger doses of chloral than are usually employed.
Paraldehyde and Chloroform.—In the experiments on blood-pressure, where the animal was under a large dose (1.2 to 1.5 c.c.) of paraldehyde, convulsions nevertheless appeared, and the mode of death was that of tutin poisoning. From such a dose of paraldehyde a rabbit may recover (Exp. 181), but the dose is a massive one, and is apparently unable to prevent the development of tutin symptoms. In these experiments, when it was
desired to keep the animal quiet it was found that chloroform-inhalation was sufficient for this purpose. The amount required seemed to be great, but there was no available means of accurately gauging the quantity. It was noted in some cases that the convulsions did not cease until the blood-pressure began to, fall, and this, taken in, conjunction with the fact that reflex excitability of the nervous system is raised, and that tutin therefore increases the effect of any reflex cardiac inhibition which may be present, points to the need of caution in its administration in cases of tutin poisoning; in fact, in one case of tutin poisoning in a rabbit, when urethane and sodium-carbonate had had no effect in easing the symptoms (Exp. 180), the administration of chloroform caused sudden cessation of the breathing, which could not be restored, although the heart continued beating and artificial respiration was carried out.
Urethane, tried in one case (Exp. 180) just mentioned, was ineffectual. A dose of 1.5 grams was given to a rabbit weighing 1.4 kilograms, and 4 mlgm. tutin was then administered. The ordinary tutin symptoms developed, though a little later than is usual with so large a dose.
Morphine.—¼ grain injected intravenously at (25) on tracing of Exp. 148 had no effect on the convulsions of a rabbit under paraldehyde. A severe convulsion followed very shortly after the injection.
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Atropine.—-1/50 grain given in the same way at (23) on tracing of Exp. 148 had no effect.
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Ilyoscine hydrobromate (Exp. 182) was found to be curiously inactive as regards rabbits. 1/100 grain in all was given to a rabbit weighing 1.5 kilograms, but apparently it had no effect at all, so its influence on tutin symptoms can be disregarded.
Bleeding is frequently used in cases of tutin poisoning, and is said to do good; but it is very difficult to judge of the value of the evidence. In the experiments on blood-pressure the animals were frequently bled to death slowly, but no cessation of the tutin spasms was observed in any case.
Restraint of the movements was tried in one experiment on a pigeon, but without influencing the result in any way.
Suspension of a rabbit under the influence of tutin by the ears or hind legs had no effect on the spasms.
Partial asphyxiation, by blocking the nostrils with a damp cloth—(26) on tracing of Exp. 148–had no effect on the convulsions; but a stream of CO2 directed against the nostrils seemed to render the breathing more regular (27).
Alkalis (Lime, &c.).—Hughes's discovery of the action of lime on tutin, although generally discredited at the time and since, may possibly be utilised to some extent in a modified form. Alkalis in general have been shown in the foregoing pages to have a very distinct action on the toxicity of tutin; thus 0.2 per cent. NaOH in five minutes at 37° to 40°C. completely destroyed the toxicity of a 0.5 per-cent. solution of tutin, and possibly the same result would follow in a shorter time and with a weaker alkali.
Since this is so, the treatment of stock poisoned by the tutu plant may be greatly improved. It is impossible to fully wash out the stomach in herbivora, so there is no way of getting rid of the leaves, &c., swallowed by the animal; but the introduction of a quantity of weak sodium-hydrate, or lime-water, or a watery suspension of magnesia simply poured down the throat, or, better, introduced by a stomach-tube, would lead to the neutralisation of the tutin present in the stomach, and so give the animal a better chance of recovery. The accumulation of gas would at the same time be
diminished by the absorption of the CO2, which no doubt forms part of the mixture of gases present, so the distension that so frequently occurs in these cases would be diminished.
The comatose condition which forms part of the symptoms of tutin poisoning suggested the possibility of some form of acid poisoning being present, and, if this be so, the injection of alkalis should prove of benefit-Sodium-carbonate was tried intravenously in one of the blood-pressure-experiments, but no apparent effect was observed. In another case sodium-carbonate was injected into the rectum of a rabbit that seemed likely to die of tutin poisoning, but here also it was ineffective. So that for the present one can only say that alkali would render unabsorbed tutin non-toxic.
19. General Summary.
1. Investigations made on the action of the pure principle tutin (C17H20O7) confirmed the results of previous observers that it is in itself sufficient to account for the main bulk, if not the whole, of the symptoms of poisoning by the tutu plant.
2. These symptoms, as they occur in cats, have been fully described, and the differences which appear in other animals noted.
3. Tutin, or its metabolic products, acts mainly on nerve-cells, producing first increased excitability and then exhaustion. It specially affects the cells of the respiratory centre, causing increased rate and depth of respiration.
4. Death may occur during the phase of increased excitability (asphyxia during convulsions) or in the phase of exhaustion. Various reflex acts—vomiting, defæcation, micturition—may occur during the stage of increased excitability. A comatose condition, possibly due to exhaustion of the cells of the cerebral cortex, is a marked feature in proportion to the strength of the dose. It deepens as death approaches. Small hæmorrhages into and congestion of the grey matter of the brain and cord are marked features in fatal cases. The Nissl granules seem unchanged when death occurs in a short time.
5. In strong solutions tutin has a slight deleterious action on tissues less highly specialised than nerve tissue—e.g., ciliated epithelium and muscle. It retards the growth of some forms of bacteria, and injuriously affects paramœcia and other low forms of life in relatively strong solutions.
6. The symptoms of poisoning by tutin are in a general way similar in widely different forms of life (flies, pigeons, cats, &c.), and can all be referred to an action on nerve-cells.
7. The minimum lethal dose in milligrams per kilo of body-weight for different classes of animals is as follows:—
|Hypodermically. Mlgm.||Per Os. Mlgm.||Immersion in. Mlgm.|
|Rabbits||" 2.5||About 6|
|Birds||Less than 5||Between 10 and 10.25|
|Lizards||Between 3 and 4|
|Frogs||" 10 and 10.25|
8. The effects on the various systems can all be referred to the influence on their nerve mechanisms—e.g.: Alimentary system, salivation, vomiting (on hypodermic injection). The circulatory system is not injuriously affected
by tutin. Cardiac inhibition does not occur, and the heart beats forcibly up to the time of death. Respiration is quickened and deepened. The pupil is not affected by local application, but dilates during the tutin, fit. General metabolism is not affected.
9. From experiments on birds and rabbits, some slight “degree of tolerance seems to be acquired. The natural relative immunity of birds is discussed.
10. Accumulation of the drug, or of its effects, may occur. Thus a guinea-pig was found to be unable to dispose of 2 mlgm. per kilo per os administered every second day.
11. The toxic action of tutin was compared with that of other members of the picrotoxin group. It was found to be, more toxic, than the sample of picrotoxin employed. The action of coriamyrtin was found to be very similar to that of tutin.
12. Attempts were made to antagonize the action, of tutin with chloral- hydrate and other drugs, with a slight degree of success.
Attention is drawn to the powerful action of weak alkalis on tutin. The toxic power of tutin is completely destroyed by 0.2 per cent. sodium-hydrate acting upon it for five minutes at 37° C., and possibly the action of weaker alkalis—e.g., lime and magnesia—would be equally destructive.
The suggestion is made that weak alkali should be used to destroy the tutin in the stomach in the case of stock poisoned by eating the tutu plant.
I may state here that the bulk of the work was done in the physiological laboratory of the Otago University, and I gladly take the opportunity of acknowledging my great indebtedness to Professor Malcolm, both for this privilege and for the essential aid he has afforded me by advice and criticism. I wish also to acknowledge my obligation to Mr. Aston for supplies of tutin and coriamyrtin, and for several references; to Dr. Hocken, for the use of his invaluable library; to Professor Benham, for the identification of specimens; to Mr. Deans, of the Acclimatisation Society, for supplies of trout-fry; and to Nurse Stronach, of the Infectious Diseases Hospital, Lake Logan, for supplies of minnows.
|P.M.||Cat (female). Weight, 3.28 kilograms|
|5.25.||Gave 33.4 minims of a 0.5-per-cent, solution of tutin in 0.75 per cent. saline (3 mlgm. per kilo body-weight), under skin of back.|
|5.29.||Respiration rapid (48 to the quarter-minute); hypersalivation; keeps mouth agape; panting vigorously.|
|5.32.||Lying on bottom of cage panting, respirations audible; mouth open; thick ropy saliva pouring from mouth; whole body shaking with respiratory effort; pupils moderate.|
|5.35.||Becoming restless; walks cautiously about cage.|
|5.36 ½.||Suddenly seized with violent convulsion; tonic spasm affecting all muscles, lasting 30 seconds, succeeded by clonic spasms; pupils widely, dilated; defæcation; micturition; nose cyanosed; unconscious, “conjunctival and ear reflex absent.|
|5.44.||Almost constant convulsion since last note; now becoming exhausted; respiration slow, irregular, spasmodic; pupils dilate during convulsions, contract in intervals.|
|5.45.||Respiration infrequent and jerky; fits in abeyance; lies full stretched on side; twitching of individual muscles.|
|5.47.||Fit after three minutes' interval; tonic spasm not succeeded by clonic.|
|5.49.||Occasional gasping respirations; writhing-movements of body.|
|5.51.||Clonic spasm, with a few involuntary cries.|
|5.53 ½.||Clonic spasm.|
|5.54.||Mistaken for dead, but still respiring feebly, and at rare intervals.|
|5.56.||Respiration ceased; heart inaudible with stethoscope; pupils dilated; dead.|
Temperature in rectum, 102° Fahr:
P.M.—Rigor mortis extremely well marked; no sign of fluid at point of injection; blood very dark and fluid.
Abdomen: Right horn of uterus contains one fœtus nearly full size, which looks as if it had died in spasm; one hind leg twisted over the other; right paw behind right ear; claws extruded.
Gall-bladder contained bile. No obvious abnormalities.
Thorax: Great veins and right side of heart distended with blood; left side contained some dark blood; lungs showed small hæmorrhages, as also did the thymus.
Brain and cord: The membranes seemed somewhat injected; grey matter of cord appeared distinctly pinkish to the naked eye.
Microscopic: The cord and medulla were hardened in 8 per cent, formol, and carried through into paraffin, and sections cut. The sections were stained by Nissl's method with toluidin blue. On examination the granules were found to be present and normal in appearance. A marked feature was the congestion of the grey matter. Capillaries can be seen close up to the nerve-cells quite full of corpuscles. At other places the collections of corpuscles suggest that minute hæmorrhages had occurred. The cord, medulla, and cerebrum show these appearances, but the congestion of the medulla is the most marked.
|P.M.||Cat (female). Weight, 3-50 kilograms.|
|5.0.||Gave 23.8 minims of a 0.5-per cent. solution of tutin in 0.75 per cent. saline (2 mlgm. per kilo body-weight), under skin of back.|
|5.1.||Gave a deep sigh.|
|5.6.||Lying in a normal attitude on bottom of cage; looks sleepy; respirations, 21 to the half-minute; pupils moderate.|
|5.10.||Still lying down; looks sleepy, and is tremulous.|
|5.14.||Opening mouth wide; cries loudly; respirations panting, 56 to the quarter-minute; has got up; salivating.|
|5.19.||Lying down; mouth wide open; panting; salivating.|
|5.21.||Slight twitching of muscles of face; cries at intervals; pupils moderate.|
|5.22.||Twitching of head.|
|5.23.||Twitching of head, followed by loud cries, as if alarmed, without knowing at what.|
|5.24.||Defæcation, large quantity of solid fæces; twitching of head.|
|5.25.||More marked twitching of head, extending to shoulders and fore paws; twitching is now almost without interval, and rapidly becoming more severe; pupils dilated, but not full; respirations irregular, suggestive of Cheyne-Stokes.|
|5.29.||General convulsion, pupils widely dilated, tonic stage lasting 30 seconds.|
|5.40.||Has continued lying on side since first convulsion; is unable to see; conjunctival reflex sluggish; convulsions have continued with intervals of only two or three seconds since last note; seems unconscious; pupils contract in intervals.|
|5.45.||Becoming exhausted, convulsions becoming feebler and rarer; respirations slow, irregular, gasping.|
|5.51.||With a final spasm and choking involuntary cry, died.|
|P.M.||Cat(female). Weight, 2-394 kilograms.|
|2.32.||Gave 5.9 minims of a 0.5-per-cent. solution tutin in normal saline (0.75 mlgm. per kilo body-weight), under skin of back.|
|2.55.||Sitting purring; pupils moderate; keeps swallowing, as if swallowing saliva; respiration, 48 per minute; “miamung” plantively; made water; defeacated (diarrhœa and flatulence), did not finish act, but walked away with discharge still pouring from anus; looks sick.|
|3.0.||Keeps mouth open; panting respiration; salivation; vomited, vomit shot out without retching.|
|3.3.||Straining at stool and “miauing”; looks ill and miserable, and disinterested in its surroundings.|
|3.5.||Begun to purr; has laid down curled up, as if to sleep.|
|3.7.||Has got up; sitting on haunches purring; twitching of muscles of face; still swallowing saliva.|
|3.9.||Lies down again; twitching of face getting more marked, and extending to shoulders; each attack of twitching followed by loud “miauing,” and, as it gets more marked, by growling; pupils dilate with each attack.|
|3.13.||General convulsion lasting one minute; saliva pouring from mouth; loud voluntary crying after convulsion; conjunctival reflex present, pupils widely dilated; can see; respiration laboured; keeps lying on side.|
|3.19.||Stands up and looks round cage; breathing easier; pupils not so large.|
|3.21.||Walked to another corner of the cage, and sat down; looks frightened and uneasy; has been no twitching for some time; “breathing quite easy, and only a little hurried; salivation seems to have ceased; constant loud crying; pupils moderate.|
|3.32.||Has been quiet since last note; looks as if it might recover.|
|3.34.||Now crying loudly, and showing slight twitching of head.|
|3.35.||Twitching getting more severe; loud prolonged plaintive cries; sitting up on haunches, looking round.|
|3.36.||Walks across cage with cautious unsteady steps.|
|3.40.||Twitching getting much more marked and more extensive, as if another general convulsion were about to occur; loud crying; whole body tremulous; respiration hurried and exaggerated, panting, with mouth open; put its head into the drinking-bowl, but did not lap; is restless.|
|3.42.||Most severe general convulsion, lasting 4 ½ minutes, then a long-drawn inspiration.|
|3.50.||Convulsions have continued almost without interval, but now becoming less severe; much frothing at mouth; gave a cry in an interval.|
|3.51.||Choky cries occur in the intervals between the convulsions; respirations irregular and gasping.|
|3.55.||A longer interval, with loud crying; respirations in the interval, 14 to the quarter-minute.|
|4.2||Has been in almost continuous convulsive movement, with comparatively quiescent intervals of only a few seconds, since 3.43; in a longer interval than usual attempted to rise, but knocked down again by a convulsion.|
|4.4.||Seizures shorter; intervals longer, about 20 seconds to 5 seconds.|
|4.6.||After a more severe convulsion than usual respirations are now very slow, irregular, and gasping.|
|4.9.||No convulsions since last note; respirations more rapid; running or swimming movement of limbs; attempts to rise, but knocked down by a convulsion.|
|4.15.||Gave a voluntary cry.|
|4.20.||Continues in constant more or less voluntary movement. Has been no convulsive seizure since 4.9 till now.|
|4.21.||No respiratory or other movement since last fit. Is dead.|
|P.M.||Cat. Weight, 2.686 kilograms.|
|3.46.||3.4 minims of a 0.5-per-cent. solution tutin (0.375 mlgm. per kilo); injected under skin of back.|
|3.58.||Defæcated, hard motion, covered it up; beginning to swallow; does not look so bright as it did; is quiet; no change in pupils; breathing more marked; beginning to look sleepy.|
|4.2.||Swallowing repeatedly; looks very sleepy.|
|4.10.||Opening mouth and panting; vomited freely and forcibly, a large quantity of stomach-contents being ejected; micturated.|
|4.20.||Twitching of eyelids and ears.|
|4.30.||Has shown twitching of face, head, and neck at short intervals; “miaus” occasionally.|
|4.40.||Twitching becoming exaggerated, involving shoulders, fore limbs, and back.|
|4.43.||General convulsions, severe.|
|4.47.||Gets up after convulsive movements have gradually subsided.|
|4.50.||Looks very ill; sitting on haunches; tremulous breathing exaggerated; answers when spoken to; twitching beginning again; salivating.|
|4.56.||Respirations 48 to the quarter-minute; lying quiet.|
|5.15.||Twitching getting more marked; cries after each attack.|
|5.21.||Twitching involving more muscles.|
|5.30.||Twitching getting very marked; at each attack animal almost springs into the air; loud crying; breathing very exaggerated; opens mouth occasionally, and pants for a few seconds.|
|5.36.||Up again; takes no notice when called; lying panting, with mouth open.|
|6.0.||Has continued twitching every few minutes since last note; now getting very marked.|
|6.5.||Severe general convulsion, lasting 1 minute.|
|6.6.||Rose again; pupils widely dilated when convulsed, diminish in size in the intervals.|
|7.15.||The cat has not been seen since last note till now; it is sitting on its haunches. tremulous, “miaus” when approached.|
|8.30.||Sitting quietly; easily startled.|
|9.30.||Apparently recovered; no symptoms noticed. Next day, 9 a.m. Seems quite well.|
|A.M.||Rabbit. Weight, 0.729 kilograms.|
|10.43.||Gave 5 minims of a 0-5-per-cent. solution tutin (2 mlgm. per kilo), hypodermically.|
|11.15.||No apparent change.|
|12.0.||Standing in middle of cage; does not appear at ease.|
|12.10.||Animal near front of cage; semi-dazed; starts when disturbed.|
|1.10.||Much the same, quiet and dazed-looking.|
|3.0.||Gave some food.|
|5.0.||Food untouched, but no obvious symptoms.|
|Next day. Quite well.|
|A.M.||Rabbit. Weight, 962 grams.|
|9.56.||Gave 8.2 minims of a 0.5-per-cent. solution of tutin in normal saline (2.5 mlgm. per kilo).|
|11.0.||Noticed to be in convulsions; last seen about 10.30, was then quiescent.|
|11.10.||Sitting up; dazed-looking.|
|11.11.||Another convulsion began, fell on side, clonic spasms; head bent back, ears twitching.|
|11.17.||Still in convulsions.|
|11.02.||Lying on side, head bent back; running-movements.|
|A.M.||Rabbit. Weight, 788 grams.|
|10.4.||Gave 8 minims of a 0.5-per-cent. solution tutin (3 mlgm. per kilo).|
|10.50.||Slow, deliberate winking.|
|10.55.||Lying on right side; breathing laboured.|
|11.41||In clonic convulsions, with movements of jaw and exposure of teeth.|
|11.15.||Still in almost constant convulsion.|
|A.M.||Rabbit. Weight, 1.507 kilograms.|
|5.12.||Gave 38.4 minims of a 0.5-per-cent. solution tutin (7.5 mlgm. per kilo), per os. Was not watched constantly, but at—|
|5.45||Was found to show great rapidity of respirations; alteration in gait—e.g., moved forward with difficulty, with body elongated, and abdomen trailing on floor; then lay on abdomen, with hind legs projecting behind, twitching of eyes and ears.|
|6.5.||Was found in severe general convulsions, head retracted, tonic spasms, well marked; running-movement of legs between fits; was held up by the ears, but this did not influence the convulsions, and no influence when suspended by hind legs.|
|6.25.||Constant movements, either convulsive seizures in which the head is always firmly retracted, or running-movements; is quite unconscious; eyes wide open, and, as it lies on its side, rubs the open lower eye into the sawdust during seizures.|
|P.M.||Rabbit. Weight, 1.16 kilograms.|
|6.0.||Gave 6 mlgm. per kilo, by mouth.|
|7.25.||Respirations seem more rapid than usual.|
|Next day, 9 a.m. Dead. Rigid, in oposthotonic posture. Has evidently died from tutin.|
|P.M.||Rabbit. Weight, 1.069 kilograms.|
|3.6.||Gave 17.5 minims of a 0.5-per-cent. solution tutin (5 mlgm. per kilo), per os.|
|3.45.||Has displayed no symptoms.|
|Next day. Quite-normal.|
Exp. 11, 12.
Two guinea-pigs, A and B. Weight of A, 624 grains (Exp. 11); weight of B, 684 grams (Exp. 12). To A, gave 2 mlgm. per kilo body-weight; to B, gave 3, mlgm. per kilo body-weight.
|3.20.||Gave to A 4.2 minims of a 0.5-per-cent. solution tutin, under skin of back.|
|3.40.||Gave to B 7 minims of a 0.5-per-cent. solution tutin, under skin of back. Both pigs were placed in the same cage, but B attacked A so viciously that it was necessary to separate them.|
|3.50.||A seems tremulous.|
|3.55.||A obviously affected, twitching and starting.|
|4.0.||B affected in the same way.|
|4.5.||Both in violent and continuous convulsions.|
|4.10.||B removed from cage, and 17 minims of a 0.05-per-cent. solution of chloral-hydrate injected (equal to 0.011 grams per kilo).|
|4.15.||B dead; killed in 35 minutes. Mouth and nostrils found blocked with sawdust from the bottom of the cage.|
|4.15.||A still in constant convulsions; becoming feebler.|
|4.30.||A dead; has continued in constant movement since convulsions appeared; killed in 70 minutes.|
Result.—A killed in 70 minutes by 2 mlgm. per kilo; B in 35 minutes by 3 mlgm. per kilo.
Guinea-pig. Weight, 751 grams.
|Feb. 13.||Gave 4 minims of 0.5-per-cent. solution tutin (1.5 mlgm. per kilo), per os.|
|" 17.||Weight, 768 grams. Gave 4 minims of 0.5-per-cent. solution, per os.|
|" 19.||" 824 grams. Gave 4 minims of 0.5-per-cent. solution, per os.|
|" 20.||Has displayed no symptoms.|
|" 22.||Weight, 809 grams. Gave 4 minims of 0.5-per-cent, solution, per os.|
|" 24.||" 823 grams. Gave 5.5 minims of 0.5-per-cent. solution tutin (2 mlgm. per kilo), per os.|
|" 25.||No symptoms.|
|" 26.||Weight, 767 grams. Gave 5.2 minims of a 0.5-per-cent. solution (2 mlgm. per kilo), per os.|
|" 27.||Found dead in the morning.|
Result.—Was able to eliminate 1.5 mlgm. per kilo given, every second day, but unable to eliminate 2 mlgm. per kilo in two days.
In this experiment the influence of the prolonged administration of tutin on the weight of the animal was tested by keeping a second guinea-pig under similar conditions as a control. The weights of the two animals as taken during the course of the experiment were as follows:—
|A (Control). Grams||B. Grams|
Result.—Control gained 38 grams; tutin animal gained 16 gram.
|A.M.||Guinea-pig. Weight, 11b. Fed up to this morning.|
|10.53.||Gave 5 minims of 0.5-per-cent. tutin; injected into mouth, and fluid was readily swallowed.|
|10.55.||Animal seems quite as usual, though easily frightened.|
|11.10.||Still same; trying to chew straw in cage.|
|11.20.||Gave it some wet green grass, which it ate greedily.|
|11.45.||Quiescent; has not eaten much grass; apparently normal.|
|12.5.||Seems still quite normal. Gave other 5 minims in same way (about 3-5 mlgm. per kilo).|
|12.20.||Stretching-movements (? normal).|
|12.33.||Sudden onset of symptoms; convulsive movements; stood on hind legs till fell over backwards; rushed several times round cage; then tonic spasm, lying on side; respiratory movement increased; twitching of ears and fore part of body. Then became quiescent, lying still on right side.|
|12.40.||Resumed usual sitting-position.|
|12.50.||Moved slowly round several times against direction, of a clock.|
|1.3.||Another fit began, with twitching of head backwards and upwards; then running-movements of limbs. It ran to front of cage, got its nose into one of the meshes, and tried, as it were, to run forward rapidly; then ran towards another corner, but fell on its side, and went into a clonic spasm, moving its fore and hind legs vigorously; then came a more tonic spasm of whole body; fine tremors; swallowing and gasping movements; mouth open. By 1.7 it became quiescent again.|
|1.10.||Another fit; lying on side and working its fore and hind legs; head bent back on body, but whole body not rigid.|
|1.15.||Still same, but movements slower.|
|1.50.||Movements continue practically without cessation.|
|2.0.||Same, slight cries.|
|2.15.||Movements less frequent.|
|2.30.||Rigor mortis seems to be setting in already.|
P.M. (3.30).—Rigor well marked; abdomen opened; some grass observed in stomach and intestines, but no great accumulation of gas.
|Jan.14.||Gave 2 mlgm. tutin per kilo body-weight, per os.|
|" 15.||No effect, so gave 4 mlgm. per kilo, per os.|
|" 18.||No effect, so gave 6 mlgm. per kilo, per os.|
|" 19.||Bird seemed dull and heavy for a few hours after the dose; took no food, and moped in a corner, with feathers puffed out. Is quite well to-day.|
|" 20.||Reweigned; same weight. Gave 8 mlgm. per kilo, per os.|
|" 21.||A repetition of symptoms noted previously. Is quite well to-day, so gave 10 mlgm. per kilo, per os.|
|" 22.||Same symptoms as before. Quite well to-day, so gave 12 mlgm. per kilo, per os.|
|" 23.||Same symptoms as before. Quite well to-day, so gave 16 mlgm. per kilo, per os.|
|" 24.||Same symptoms as before, but quite well to-day; so at 12 noon gave 20 mlgm. per kilo, per os, on an empty crop. At 12.45 p.m. bird found lying on back in convulsions; marked retraction of head, and constant movements of feet and flapping of wings. At 12.50 p.m. movements becoming feebler and slower. At 1 p.m. dead; weight, 345 grams, an increase of 31 grams.|
P.M.—Crop noted to be empty; nothing abnormal detected.
Result.—16 mlgm. per kilo non-lethal; 20 mlgm. per kilo killed in one hour.
|A.M.||Pigeon. Weight, 319 grams.|
|10.6.||Gave 18.3 minims of 0.5-per-cent. solution tutin in normal saline (17 mlgm. per kilo body-weight), per os.|
|10.8.||First appearance of symptoms, tremulousness and jerking of head; sudden slight expansive movements of the wings.|
|10.10.||General convulsions; marked opisthotonus; bird turning over and over, head firmly retracted on back, wings/widely extended, general convulsive movements.|
|10.13.||Has been no cessation of convulsive movements since onset; now becoming more feeble.|
|10.21.||Movements very feeble; bird apparently been unconscious since onset.|
|10.22.||All movements ceased.|
P.M.—Crop opened; found empty, except for a small amount of grumous fluid containing small yellow particles. Sour smell, and distinct acid reaction. No abnormality observed.
Result.—Death in 16 minutes from 17 mlgm. per kilo.
|Feb. 7.||At 6 p.m. gave 2 mlgm. of a 0.5-per-cent. solution tutin (about 1.5 mlgm. per kilo), per os. At 7.5 p.m., apparently normal. At 10.45 p.m., same.|
|" 8.||Quite normal. Gave 3 mlgm. of a 0.5-per-cent. solution tutin (about 2 mlgm. per kilo), per os.|
|" 10.||At 6 p.m. pigeon weighed 396 grams. Gave 5.3 minims of a 0.5-per-cent. solution tutin (4 mlgm. per kilo), per os.|
|" 14.||Weight, 408 grams. Gave 9.7 minims, of a 0.5-per-cent. solution (9 mlgm. per kilo), per os.|
|" 17.||At 6.3 p.m. weighed 388 grams. Gave 11.8 minims of 0.5-per-cent. solution tutin (9 mlgm. per kilo), per os.|
|"19.||Weight, 342 grams. Gave 10.8 minims of 0.5-per-cent., solution tutin (9.5 mlgm. per kilo), per os.|
|" 24.||Weight, 343 grams. Gave 11.6 minims of 0.5-per-cent. solution tutin (10 mlgm. per kilo), per os.|
|" 26.||At 6 p.m. weighed 370 grams. Gave 15.4 minims of a 0.5-per-cent. solution tutin (12 mlgm. per kilo), per os. At 6.20 p.m. squatting on floor of cage; very tremulous about head and neck; blinking repeatedly.|
|" 27.||Pound dead in the morning.|
Result—No tolerance developed. Death from a dose of 12 mlgm. per kilo.
|Jan. 28.||Pigeon. Weight, 357 grams.|
|5.45.||Gave 18.2 minims of a 0.5-per-cent. solution tutin in normal saline (15 mlgm. per kilo body-weight), per os.|
|7 30.||Beyond some dullness and heaviness, no symptoms.|
|Next day. Quite normal.|
Result.—Recovery from 15 mlgm. per kilo.
|P.M.||Pigeon. Weight, 374 grams.|
|6.7.||Intended to give 20-3 minims of a 0.5-per-cent. solution tutin (16 mlgm. per kilo body-weight), but a few, drops were lost, so gave 5 minims more.|
|6.10.||General convulsions; marked extension of wings and bending-backwards of the body, so that the bird rests on its tail.|
|6.12.||Lying on back, in constant convulsive movement.|
P.M.—Crop half-full. No abnormality detected.
Result.—Death in 9 minutes from a dose of about 16 mlgm. per kilo.
|P.M.||Pigeon Weight, 319 grams|
|5.6.||Gave 17-3 minims of a 0.5-per-cent. solution tutin (16 mlgm. per kilo body-weight), per os.|
|5.7.||Appearance of symptoms.|
|5.7.||½. Clonic convulsions began.|
|5.11.||Movements ceased; pupils dilated; dead.|
P.M.—Crop full of half-digested food. No abnormalities observed.
Result.—Death in 5 minutes from a dose of 16 mlgm. per kilo.
|P.M.||Pigeon. Same as used in Exp. 14. Weight, 345 grams.|
|6.14.||Gave 16.8 minima of a 0.5-per-cent. solution tutin (15 mlgm. per kilo body-weight), per os. Had received no food since the day before.|
|5.20.||Seems quite normal; is preening its feathers.|
|5.21.||Attempting to vomit.|
|5.24.||Retching; drowsy-looking; tremulous about head.|
|5.25.||Squatted down on bottom of cage.|
|5.26.||Very tremulous about head and neck; dazed and sleepy looking; retching every half-minute or so.|
|5.29.||Twitching of muscles of neck; head being drawn back with sharp jerks.|
|5.36.||Slight convulsions affecting the fore part of the body; wings rigidly extended, and marked clonic movements of tail and neck; remains standing between the attacks; blinks its eyes heavily, as though only keeping awake with the greatest effort; pupils contracted.|
|5.40.||A more severe convulsion; bird thrown over on to its back, but recovered, its position again.|
|5.43.||A most severe convulsion, bird thrown over backwards; lies on back, with head bent back beneath body; all muscles in continuous clonic spasm.|
|5.49.||Convulsions have continued without intermission since last note; movements now becoming feeble.|
|5.50.||All movements ceased; dead.|
P.M.—Crop empty. No abnormality observed.
Result.—Killed in 35 minutes by a dose of 15 mlgm. per kilo.
|P.M.||Pigeon. Weight, 203 grams.|
|6.13.||Gave 10.3 minims of a 0.5-per-cent. solution tutin (15 mlgm. per kilo), per os.|
|6.20.||Lying on back, in continuous convulsive movement.|
|5.25.||Gave 10 minims of a 1-in-1 solution chloral-hydrate, equal to 0.6 gram chloral, per os.|
P.M.—Crop half-full. No abnormality observed.
Result.—Death in 72 minutes from 15 mlgm, per kilo. Life evidently prolonged by the chloral.
|P.M.||Pigeon (young). Weight, 375 grams. (Fed at 1 p.m.)|
|3.49.||Gave 15.5 minims of 0.5-per-cent. solution tutin (13 mlgm. per kilo body-weight), per os.|
|3.52.||Slight attempts at vomiting.|
|4.0.||Lying on back, in convulsions; convulsive movements continued till death.|
Result.—Death in 19 minutes from 13 mlgm. per kilo.
|P.M.||Pigeon (young). Weight, 363 grams.|
|3.46.||Gave 14.8 minims of 0.5-per-cent. solution tutin (12 mlgm. per kilo body-weight), per os.|
|3.50.||Vomiting repeatedly, bringing up quantities of grain.|
|4.8.||Is tremulous and unsteady.|
|4.14.||Slight convulsive movements, affecting the wings, and head; vomiting.|
|4.25.||Defæcated; a more severe convulsion; body bent forward till breast touches ground, head bent back, wings rigidly extended, perhaps one more than the other. The bird was here taken out of the cage and placed on the floor of the laboratory; it did not attempt to fly away, and kept pretty much to the place where it was set down; it moved away a little if approached. On the whole, seemed more acute mentally than birds that had received a larger dose.|
|4.28.||Convulsion, but still able to keep its feet; pupils widely dilated during convulsion.|
|4.38.||Vomiting and convulsions.|
|4.39.||Convulsion, still affecting chiefly the fore part of the body.|
|4.42.||Vomiting and convulsions; pupil continuously dilated|
|4.46.||Most severe convulsive attack yet, tail involved, and clomic as well as tonic spasm present. The bird was thrown down, but recovered its feet again.|
|5.10.||Since last note have been frequent convulsions, affecting the neck; wings, and tail, but the bird has kept its feet. They are getting more severe and last longer.|
|6.12.||Most severe convulsive seizure, bird tumbling and somersaulting about the floor of the laboratory.|
|5.14.||Lying on back; all the muscles in constant clonic spasm.|
|5.16.||Short interval, during which voluntary squeaking.|
|5.20.||Lying on back; convulsions continue; legs as much affected as wings.|
|5.42.||Has been lying on back in constant movement since 5.16.|
|5.51.||Movements becoming more feeble; they never entirely disappear, but increase in severity at intervals. In the intervals, gasping inspirations.|
|6.2.||All movement ceased; dead.|
Result.—Death in 136 minutes from a dose of 12 mlgm. per-kilo.
|P.M.||Pigeon (adult). Weight, 386 grams.|
|4.45.||Gave 15.7 minims of 0.5-per-cent. solution tutin (12 mlgm. per kilo), per os.|
|5.30.||Lying on back, in continuous convulsive movement.|
|5.31.||All movement ceased; dead.|
Result—Death in 46 minutes from a dose of 12 mlgm. per kilo.
|P.M.||Pigeon (adult). Weight, 305 grams.|
|5.52.||Gave 9.3 minims of a 0.5-per-cent. solution tutin (9 mlgm. per kilo), per os.|
|Next day, 4 p.m. Quite normal. Displayed no marked symptoms.|
Result.—Recovery from a dose of 9 mlgm. per kilo.
|P.M.||Pigeon (adult). Weight, 365 grams.|
|4.30.||Gave 11.7 minims of a 0.5-per-cent. solution tutin (9.5 mlgm. per kilo), per os.|
|Next day, 4.30 p.m. Quite normal. Has displayed no marked symptoms.|
Result.—Recovery from a dose of 9.5 mlgm. per kilo.
|P.M.||Pigeon (young). Weight, 330 grams.|
|4.45.||Gave 11.2 minims of a 0.5-per-cent. solution tutin (10 mlgm. per kilo), per os.|
|5.10.||Lying on back in convulsions.|
Result.—Death in 45 minutes from a dose of 10 mlgm. per kilo.
|P.M.||Pigeon (adult). Weight, 368 grams.|
|4.56.||Gave 10.6 minims of a 5-per-cent. solution tutin (10 mlgm. per kilo), per os.|
|5.21.||Slight convulsive movements of wings.|
Result.—Recovered from a dose of 10 mlgm. per kilo.
|P.M.||Pigeon. Weight, 315 grams.|
|2.50.||Gave 11.2 minims of a 0.5-per-cent. solution tutin (10.25 mlgm. per kilo), per os.|
|3.18.||Tremulous; jerking of head; restless; blinking.|
|3.20.||Tonic spasm of wings; paroxysms of difficult breathing, during which bird turns round and round in one place.|
|3.48.||General convulsion, lasting 30 seconds, in which bird tumbles about in every direction.|
|3.51.||Lying on side, trembling and breathing very rapidly; pupils normal.|
|3.53.||General convulsion; pupils wide.|
|3.56.||Recovered ‘upright position, and sat for a few seconds on its tail; was then seized with severe convulsions, which continued without intermission till death.|
Exps. 31, 32. (See text.)
|P.M.||Lizard (Lygosoma moco). Weight, 4 grams.|
|5.15.||Gave 3.4 minims of a 0.01-per-cent. solution tutin (5 mlgm. per kilo), under the skin of the abdomen.|
|7.10.||Quiet when first seen; then took fits of abnormal activity, contorting itself, holding its fore limbs wide stretched, and resting on belly.|
|7.55.||Lying quiet, with mouth partially open.|
|8.5.||Seems quite dead; mouth still more widely open.|
|P.M.||Lizard. Weight, 5 grams.|
|3.0.||Gave 2.5 minims of a 0.01-per-cent. solution (3 mlgm. per kilo), under the skin of the abdomen. The breathing became exaggerated almost immediately, and the animal puffed itself up as a frog does.|
|7.30.||Nothing remarkable has been noticed.|
|Next day, 9 a.m. Apparently normal.|
|P.M.||Lizard. Weight, 7 grams.|
|4.35.||Gave 5 minims of a 0.01-per-cent. solution of tutin (4 mlgm. per kilo), hypodermically.|
|7.20.||Remarkable convulsive effects, opisthotonus and twisting into a ball with tail up to mouth, then clonic spasms of limbs; after this was quiet for a time, and then movements began again; lies on back biting at its own tail and hind limbs.|
|7.45.||Has been quiescent for last five minutes; now puffing itself up;lying in normal position.|
|7.58.||Another severe fit coming on suddenly; animal twists rapidly into every possible attitude.|
|8.2.||Still in constant movement, now more often on its back.|
|8.25.||Another period of restlessness.|
|8.35.||Has quiet intervals.|
|9.15.||Seems quite dead, but gave reflex response from limbs, and did not move after that.|
Exps. 36, 37, 38.
|4.26.||Gave to frog (1), 4.1 minims of 0.01-per-cent. solution tutin (1 mlgm. per kilo); to frog (2), 3.4 minims 0.025-per-cent. solution tutin (2 mlgm. per kilo); to frog (3), 6.8 minims 0.025-per-cent. solution tutin (3 mlgm. per, kilo). In each case the solution was injected into the abdominal cavity, and the frogs placed under bell-jars on the laboratory-table.|
|7.30.||Frog (3) seems affected; crouches in a sitting-position, and is blown out with air. No apparent abnormality in frogs (1) and (2).|
|10.0.||No sign of convulsion in any when stimulated. Frog (3) seems more excitable than the others.|
|8.55.||All three appear to be affected; are sluggish, and lying prone on their bellies, with legs extended. Frog (2) is almost dead.|
|10.0.||Frog (2) dead (2 mlgm. per kilo); died between 9 and 10 a.m.|
|Jan. 18.||Frog (1) found dead in the morning (1 mlgm. per kilo); weight, 18 grams, a loss of 9 grams. Frog (3) (3 mlgm. per kilo) is still alive, and seems quite normal.|
No sign of convulsion noticed in these frogs. The two that died seemed extremely thin and emaciated. Frog (1) looked very much thinner on the 17th than, on the two days before. The weather was cold on the 16th. On the 17th the temperature was 12° C. in the morning, but this does not explain why frog (3); with 3 mlgm. per kilo, seems unaffected. Loss of weight might be due to drying; but the air has not been dry. The weather is cold and wet.
Exps. 39, 40, 41.
|4.40.||Gave to frog (1), 2-6 minims of a 0.01-per-cent, solution tutin (1 mlgm. per kilo); to frog (2), 7-5 minims of a 0.01-per-cent. solution tutin (2 mlgm. per kilo); to frog (3), 5-7 minims of a 0.025-per-cent. solution tutin (3 mlgm. per kilo). In each case the solution was injected into the abdominal cavity, and the frogs placed under bell-jar on the laboratory-table.|
|10.0.||No change observed.|
|Jan. 17.||No change observed.|
|Jan. 18.||Frog (2) was found dead in the morning; weight, 18 grams. Weather cold.|
Frogs (1) and (3) were apparently normal.
In the two experiments above, the number of minims in 1 c.c. was taken as 15; in the subsequent experiments as 17. The doses per kilo body-weight above are therefore smaller than as stated.
Exps. 42, 43.
|5.0.||Gave to frog (1), 4 minims of a 0.05-per-cent. solution tutin (4 mlgm. per kilo).|
|5.15.||Gave to frog (2), 7 minims of a 0.05-per-cent. solution tutin (5 mlgm. per kilo).In each case the solution was injected under the skin of the back, and the frogs were placed under bell-jars, with moist grass and earth, in a cool dark cellar.|
|7.30.||Frog (1) gave a long cry when touched; seems very excitable. Frog (2) apparently normal.|
|10.30.||Could detect nothing abnormal in either.|
|9.30.||Both apparently well.|
|2.30.||Both quite normal.|
|Jan. 25.||Quite normal.|
|3.0.||Gave 9.5 minims of a 0.05-per-cent. solution tutin (7 mlgm. per kilo), injected - under the skin of the back. The frog was placed under a bell-jar, with wet grass, in a cool cellar.|
|6.30.||No obvious effect.|
|10.30.||No obvious effect.|
|9.0.||No obvious effect.|
|4.30.||No obvious effect.|
|9.30.||Apparently quite normal.|
|3.35,||Gave 5*5 minims of a 0.05-per-cent. solution tutin (9 mlgm. per kilo), injected under the skin of the back. It was placed in a cellar, with moist grass and earth.|
|6.10.||Not normal; squats flat on belly; muscular weakness of hind legs; respiration exaggerated.|
|7.30.||Hind limbs show spastic spasms; unable to progress; lies sprawling on belly.|
|10.45.||Seems much improved; aits in normal position; hops away unsteadily when touched.|
|Jan. 30.||Seems weak, but in other respects is quite normal.|
|5.33.||Gave 13 minima of a 0.05-per-cent. solution tutin (10 mlgm. per kilo), injected under the skin of the back. It was placed in a cool cellar, with moist grass.|
|9.40.||Frog found in severe spasm, chiefly affecting the hind limbs, which are fully extended; is very excitable; gave cries when touched, and climbed to the top of the bell-jar; no muscular weakness manifest.|
|9.0.||Very lively and excitable; no distinct convulsions when touched; is still noisy.|
|2.30.||Just dying; mouth open; could not obtain reflexes.|
|Jan. 31.||Gave 11 minims of a 0.05-per-cent. solution tutin (9 mlgm. per kilo), injected under the skin of the back. It was placed in a cool cellar, with moist grass.|
|Feb. 3.||Quite normal.|
|6.15.||Gave 10.5 minima of a 0.05-per-cent. solution tutin (10 mlgm. per kilo), injected under the skin of the back. As before, it was placed in favourable surroundings.|
|3.45.||Seems weak, but shows no marked symptoms.|
|Feb. 5.||Apparently quite normal.|
Exps. 49, 50, 51.
Three frogs. Weights: (1),23 grams (Exp. 49); (2) 39 grama (Exp.50); (3), 30 grams
|6.20.||To frog (1) was given 7 minims of a 0.05-per-cent. solution tutin (9 mlgm. per kilo).|
|6.24.||To frog (2) was given 13.2 minims of a 0-05-per-cent. solution tutin (10 mlgm. per kilo).|
|6.31.||To frog (3) was given 11.2 minims of a 0.05-per-cent. solution tutin (11 mlgm. per kilo).|
|In each ease the solution was injected under the skin of the back, and the frogs were placed in favourable surroundings.|
|4.0.||Fiog (1) affected; lying prone on belly, with legs extended; great muscular weakness. Frog (2) presented the same appearance. Frog (3) was found dead at 9 a.m.|
|Feb. 20. Frogs (1) and (2) apparently normal.||" 21. Both frogs quite well.|
Frog. Weight, 40 grams.
|5.0.||Gave 14 minims of a 0.05-per-cent solution tutin (10-25 mlgm. per, kilo). The solution was injected under the skin of the back, and the frog placed in a cool cellar, with moist grass and earth.|
|2.0.||Sprawling on belly; hind legs extended; abdomen distended; twitching of toes; fibriliary twitching of muscles of thighs. On being handled, was seized with a convulsion, affecting chiefly the hind legs; the mouth was opened wide and kept open, the animal croaking loudly.|
Pigeon. Weight, 335 grams.
|5.30.||Gave 5.6 minims of a 0.5-per-cent. solution tutin (5 mlgm. per kilo). The injection was made under the right, wing.|
|5.45.||Very sleepy-looking; narcotic effect is most marked; tremulous about head; retching.|
|6.5.||General convulsive movements have continued since last note; are now becoming feebler; bird on its back.|
|6.12.||All movement ceased; dead.|
|Result—Death in 42 minutes from a hypodermic dose of 5 mlgm. per kilo.|
Frog. Weight, 20 grams.
|4.55.||Gave 6.8 minims of a 0.1-per-cent. solution tutin (20 mlgm. per kilo), under the skin of back.|
|5.0.||Breathing exaggerated; keeps opening mouth wide.|
|5.15.||No further symptoms; gave other 7 minims.|
|5.16.||Rapid breathing; mouth-opening movements.|
|5.26.||Gave other 7 minims.|
|6.0.||Lying prone on belly; shows occasional slight twitchings, chiefly of hind legs; if touched, is seized with convulsions, and pupils, dilate; respirations irregular and exaggerated; makes a sucking noise occasionally.|
|6.9.||Turned on back; cannot recover, and struggles violently and continuously in its efforts to do so.|
|6.11.||Most severe convulsions; mouth spasmodically opened to fullest extent; cries loudly; spasms occur every 4 or 5 seconds (This change has occurred since, and apparently as a result of, violent voluntary efforts to recover position when placed on back.) Heart can be seen beating through abdominal wall; rate, 44 per minute.|
|6.20.||Heart-movement not so visible; rate, 32 per minute. Muscles still show occasional twitches and spasms.|
|6.30.||Seems quite dead; no reflexes.|
|This frog received about 60 mlgm. per kilo.|
Frog (female). Weight of frog, 38 grams; ovaries, 57 grams.
|4.12.||Gave 1 minim saturated solution tutin, injected under skin of back.|
|4.30.||Remains crouching; if turned on back, recovers position slowly.|
|4.35.||Turned on back; recovers position with difficulty; convulsive movements fore and hind limbs; lies prone on belly; if legs extended, does not retract them; respiration feeble and irregular; convulsive movements of limbs if turned on back.|
|4.40.||Attempted to crawl, but unable to advance; pricking skin of limb, no response.|
|4.45.||Respiration imperceptible; if turned on back, recovers position with difficulty, then tonic spasm in extension and a few vigorous respirations.|
|4.55.||When turned on back, quickly recovered, and made three or four vigorous hops; crying loudly, with jaws spasmodically opened, then lay on belly, and at intervals of 10 to 15 seconds opened jaws widely and extended limbs in tonic spasm.|
|5.15.||When turned on back, lay so, occasional tonic spasm of fore limbs, irregular feeble movement of hind limbs; respiration at rare and irregular intervals—two or three respirations, and then a more or less prolonged pause; helpless, lying on belly, reflexes still present; makes no attempt to swim when placed in water; unable to rectify position in water; lying on back at bottom of basin.|
|6.7.||Under bell-jar, makes occasional kicking-out movements; still on belly, and swollen-looking.|
|7.40.||Very similar; makes less movement than before; hes in practically any position if gently handled, but reflex action still marked; pupils dilated|
|10.15.||No spontaneous movement, and no reflexes (some movements, got on turning over or letting fall, seem due to direct stimulation of muscles); pupils dilated; no respiration seen.|
|Next day, 9 a.m. Dead; stiff in position it was in for greater time after injection|
|3.0.||Gave 1 minim semi-saturated solution tutin, injected under skm.|
|3.3.||Attempting to jump, but movements already enfeebled|
|3.15.||Respiratory movement exaggerated; abdomen distended.|
|3.20||Slowly extended hind legs and advanced fore legs, and lay on belly; respiration quickened and exaggerated; occasionally raises fore part of body in spasm to full extent of fore arms, and respiration ceases.|
|3.25.||Twitching of toes.|
|3.35||Attempting to crawl flat on belly; unable to make progress; conjunctival reflex active; skm reflex sluggish; pupils dilated.|
|3.40.||Spasm of all four limbs; restless; raises body stiffly, with hind limbs extended.|
|3.45||Continuous struggling-movements; twice turned on back, and slowly and with difficulty recovered position|
|3.50.||Placed on back, unable to recover; twitching of muscles and successive tonic spasms; respiration rare and convulsive; has opened mouth spasmodically once.|
|4.10.||Reflex action abolished, except conjunctival, which is sluggish.|
|4.15.||Repeated spasmodic opening of mouth|
|4.30.||Can still see; lies on back; occasional movements of fore limbs.|
|5.5.||On letting fall, slight reflex in fore limbs; conjunctival reflex still present.|
|4.30.||One drop of a semi-saturated solution tutin in conjunctival sac.|
|4.45.||Movements of fore limbs to eyes; conjunctival irritation; restlessness; exaggerated respiration.|
|5.0.||Opisthotonic movements; weakness of hind limbs; crawls slowly; hops with difficulty.|
|7.5.||When light turned on, was found to be in a state closely resembling clonic stage of epileptic fit; this soon subsided, but could be initiated to a lesser degree by pulling animal about by hind leg. Pupils wide; respiratory movement excited and irregular.|
Frog (small). Weight, 12-5 grams.
|12.35.||Gave 1 minim of a 0.05-per-cent. tutin solution in 0.76 per cent, saline, injected under skin. Motionless, on belly.|
|12.42.||First movement, lifted head.|
|12.45.||Sprang forward; increased respiratory rate.|
|12.50.||Attempted to spring, but moved sluggishly, and did not advance; emptying of cloaca, urine, and fæces.|
|12.55.||Raised on all four limbs, and sank slowly back on belly.|
|1.0.||Raised himself on all-fours, and remained sitting up with fore limbs extended; irregular crawling-movements, chiefly in fore limbs; feeble attempts, to spring; hind limbs seem weak; pupils dilated.|
|1.10.||Restless; attempting to crawl up side of bell-jar; movements incoordinate and feeble.|
|1.43.||Sitting motionless; pupils more dilated than at beginning of experiment; can leap fairly well when toe pinched, but seems feebler, and does not resent interference so much as in a normal frog.|
|2.3.||Leaning up against side of bell-jar; left hand partially closed, as if grasping a twig; right extended against glass.|
|2.43.||Lying quiet; recovers position fairly quickly when turned on back; also shows occasional voluntary movements|
|3.13.||Same; pupils wider than previously.|
|4.13.||As before, but respirations (taken from movement of floor of mouth) 40 for one half-minute, 30 for another half-minute, and rather irregular.|
|4.50.||Lying prone on belly, with legs extended from sides; if turned on back, recovers with difficulty; able to advance across table by short halting leaps; hind legs drawn up slowly to jumping-position after each effort; no convulsive movements noted so far; seems to be a general, muscular enfeeblement, most marked m hind legs.|
|5.0.||Makes continued efforts to jump, but fails to advance, as cannot flex legs sufficiently beneath body.|
|6.0.||Lying prone on belly; turns over when placed on back, though with difficulty.|
|7.36.||Lying sprawling on belly; limbs extended; moves feebly when disturbed; pupils dilated (? due to darkness); while being observed it had something like a weak convulsive fit. It lay on belly and kicked out repeatedly but feebly with hind legs, and moved fore limbs as if attempting to swim.|
|9.35.||Sprawling on belly as before, and motionless till disturbed. Weakness more marked; cannot turn over when placed on back; makes several efforts, accompanied by deep breathing, and then lies still.|
|Next day, 9am Found dead, in same position as left in last night; pupils firmly contracted; lower limbs extended; fore limbs flexed, and digits closed, as if grasping twig; no swelling of abdomen.|
|P.M.—Gall-bladder distended; intestines distended; cloaca full; intestine, some daik grumous liquid; stomach empty, except for mucus; kidneys apparently normal; no abnormality at seat of injection; central nervous system nothing abnormal, except seems too pale, no ecchymosis; blood seems fluid, and animal as a whole seems more bloodless than normal; blood-film squeezed out of heart and tissues presents many leucocytes, possibly due to mode of obtaining it.|
|5.45.||Small trout, m 100 c.c. water, with 10 drops saturated solution of tutin (about 0.1 per cent.).|
|5.49.||Movement very excited; breathing exaggerated.|
|6.18.||Breathing more laboured.|
|6.28.||Swimming on side, near surface.|
|6.30.||Shuddering movements; spasmodic movements of gills, which then ceased. Control, in same amount of water, showed no change.|
Exps. 60 to 76.
These experiments were very like the above (see Table III).
Table, of Experiments on Fishes.
|Exp-No||Drag.||Percentage of Drug||Modification of Drug.||Volume of Flund.||Number of Fishes.||Time of Onset of Symptoms.||Result||Time of Death.|
|78||"||0.00075||200||1||5 hours||"||6⅓ "|
|79||"||0.005||200||1||5 "||"||6 "|
|81||"||0.0075||200||1||5 hours||"||6 "|
|82||"||0.005||200||1||1¼ "||"||4 "|
Control to the above fishes tinder the same conditions (200 c.c. fluid) showed symptoms of asphyxiation in 4 hours, and the fishes in tutin and picrotoxin died practically in order of their weights.
|Exp-No||Drug.||Percentage of Drug.||Modification of Drug.||Volume of Flund.||Number of Fishes.||Time of Onset of Symptoms.||Result.||Time of Death.|
|84||Tutin and chloral||0.001||1,000||3||Not observed||1 died||About 22 hours.|
|86||Tutin||0.0015||1,000||3||1 hour||"||1 5/3; hours.|
|87||"||0.00125||1,000||3||No symptoms||All lived|
|88||"||0.00175||1,000||3||4½ hours||1 dled||7 hours.|
|(1.) 2¾ hours||Died||7 hours.|
|89||"||0.003||1,000||3||(m.) 9½ "||"||12 "|
|90||"||0.002||1,000||3||No symptoms||All lived Died||4½ hours.|
|91||"||0.0035||1,000||3||(1) 4 hours (m) Slight symptoms from 7–10 hours (s) No symptoms||Recovered|
|92||"||0.0035||Digested with HCI 30 min. and then neutralised||1,000||3||(1) 3¾ hours
(m) 8⅓ "
(s.) 9 "
|Fatal to all||(1.) 7½ hours.
(m) 10–22 "
(s.) 22½ "
|93||"||0.003||Already once used, filtered||1,000||1*||4⅔ hours||Died||6⅔ hours|
|94(A)||"||0.004||Hydrolysed with H Cl 1 hour at 37° C.||1,000||3||(1.) 5 hours
(m.) 6¼ "
(s.) 8 "
|95(B)||"||0.004||Control to 94; HCl added, and immediatley neutralised||1,000||3||(l.) 3¾ "
(m.) 8 "
(s) Shight symptoms at 12 hours
|96(C)||"||0.004||Treated with 02 per cent. NaCH for 1 hour at 37° C||1,000||3||No symptoms||All lived|
|97||"||0.004||Hydrolysed with HCl 1 hour at 37°||1,000||3||(1.) 3 hours
(m.) Symptoms at 16 hours
(s) No symptoms
|About 16 hours.
" 22½ "
|98(B)||"||0.004||Control to A. HCl immedately neutralised||1,000||3||2 hours||All died||About 16 hours.|
|99(C)||"||0.004||Treated with 0.2 per cent. NaOH for I hour at 37° C||1,000||3||No symptoms||All lived|
|100||Picrotoxin||0.004||1,000||3||(1.) 3½ hours (m.) 3½ " (s.) 6½ "||Died
[Footnote] * Fish used in this experiment was Cheimarrichthys fosteri. Tutin solution, 0.003 per cent., was not fatal to three minnows.
Table of Experiments on Fishes—continued.
|Exp No.||Drug.||Percentage of Drug.||Modification of Drug.||Volume of Fluid||Number of Fishes.||Time of Onset of Symptoms.||Result.||Time of Death.|
|101||Tutin||0.005||Treated with 0 2 per cent. NaOH for 20min||CC 1,000||3||No symptoms.||All lived 25 hours|
|102||"||0.005||Treated with 0 2 per cent. NaOH for 40 mm.||1,000||3||No symptoms||All lived 25 hours|
|103||"||0.005||Acted on by living livercells for 1 hour at 37° C.||1,000||3||(1.) 2⅓ hours
(m.) 2 ⅓"
|All died||5½ hours.
|104||"||0.005||Acted on by living kidney-cells for 1 hours at 37° C.||1,000||3||(1.) 1½ hours
(m.) 2 ⅓ "
(s.) 4½ "
|105||Plcrotoxin||0.0132||1,000||3||(1.) 1½ "
(m.) 1½ "
|106||Tutin||0.005||1,000||3||(1.) 7½ "
(m.) 6½ "
(s.) 7½ "
|"||About 22¼ hrs.
23 hours. About 22¼ hrs.
|107||Picrotoxin||0.005||1,000||3||(1.) 1 ⅔ "
(m.)1 ⅔ "
(s.) 1 ⅔ "
11 ⅔ "
12 ⅔ "
|108||Tutin||0.005||Had been used once before||1,000||3||(1.) 2⅘ "
(m.) 4⅓ "
(s.) 5½ "
|12½ hours. 12½-25 hours.|
|109||"||0.005||Treated with 0 2 per cent-NaOH for 10 minutes||1,000||3||No symptoms||All lived 30 hours|
|111||"||0.005||Treated with 0 2 per cent. HCl for 1 hour at 37° C.||1,000||3||(1.) 6⅔ hours
(m.) 5⅓ "
(s.) 51/3 "
|All died||13⅚ hours.
|112||"||0.005||Treated with 0 2 per cent. NaOH for 5 minutes at 37° C.||1,000||3||No symptoms||All lived 26 hours|
Flinds 94 (A), 95 (B), and 96 (C) filtered and tried over again. (1.), (m.), (s.) = large, medium, small.
Three minnows (large, medium, and small).
|8.43.||Placed in 1,000 c.c. fluid of 0 005-per-cent. solution.|
|11.30.||Large fish on side, emitting air-bells.|
|11.50.||Large fish now upright, but keeping near surface.|
|12.10.||Large fish on its side; movements feeble. As yet the other two fish are unaffected.|
|12.49.||Large fish upright again; near surface.|
|1.5.||Large fish on side near surface. Medium fish in excited, movement near surface, emitting air-bells. Small fish excited.|
|2.15.||Large fish same; medium and small both on the side near the surface.|
|4.0.||All markedly affected, and keep more or less on side, near the surface.|
|6.20.||All swim near surface, excitable, but feeble; now swimming upright.|
|7.20.||All keep close to surface, and emit air-bells.|
|8.20.||Gill-movements have ceased in medium-sized fish; heart-movement still visible. Large and small fish both deeply affected.|
|9.15.||Large fish dead. Small fish deeply affected; swimming upright, but very feebly, near surface.|
|Next day. Small fish apparently well.|
|3.55.||Two cockles, of about the same size, were placed—one in sea-water, the other in a 0 5-per-cent. solution of tutin in sea-water. Equal volumes of fluid were used.|
|4.0.||The cockle in tutin solution has opened its shell.|
|4.10.||The cockle in sea-water has not moved. The one in tutin solution keeps opening and closing its shell at intervals of a minute or two.|
|4.15.||Portion of the one in tutin solution is now extruded from its shell; the shell every now and then shuts down sharply, but without the extruded portion being drawn back; when touched with a needle the extruded portion is withdrawn, but is pushed out again after a short interval.|
|4.25.||The one in tutin solution continues extruded,. but withdraws when the glass container is tapped with a pencil.|
|Next day, 9 p.m. The one in tutin solution is dead; throughout the day it has remained with shell open and body extruded; at noon to-day was alive, and withdrew sluggishly when touched with a needle. The one in sea-water is still alive, with the shell firmly closed; it has not been observed to open the shell throughout the experiment; on being placed in the tutin solution it opened its shell in a few minutes, and behaved as the other had done.|
|Jan. 21.||At 8 p.m. two flies—a large blowfly and an ordinary house-fly—were placed in a cage with a watch-glass containing a solution of sugar in a 0 1-per-cent. solution tutin. Both were observed to partake freely of the solution on several occasions.|
|" 22.||At 8.30 p.m. both flies are active, and apparently quite normal; they frequently feed from the watch-glass|
|" 23.||At 12.15p.m. the house-fly is dead. The blowfly is active, and apparently normal.|
|"24.||Blowfly quite well; allowed to escape.|
|Mar. 1.||About twenty house-flies were placed in each of two cages. Into one cage was put a solution of sugar in a 0 1-per-cent. solution tutin in normal saline; into the other a solution of sugar in normal saline. The flies, being thirsty, at once crowded round both watch-glasses, and partook freely|
|" 4.||All the flies are alive, and quite healthy. The flies with the tutin solution do not feed so readily as those with the normal saline; they frequently approach the watch-glass, merely taste the solution, and retire; now and again one will remain feeding.|
|" 7.||Flies still all alive, and quite healthy; during the week the solution has dried up once or twice, and more 0 1-per-cent. solution tutin has been added. Despite the concentration by evaporation, they have displayed no symptoms.|
|6.40||A few drops of 0.5-per-cent. solution tutin in distilled water added to the watch-glass, which, had become dry.|
|7.0.||On returning, found four flies on their backs, but not dead; moving their legs occasionally. Three other flies were affected. Every now and then wings violently buzzed for a second or two, lifting flies on to tip-toes. They walk rather mcoordinately.|
|7.5.||One of the affected flies spinning round and round on its back, wings buzzing continuously; movement ceases for a short interval, and is then repeated.|
|7.8.||Both the other flies similarly affected.|
|7.15.||All three are now lying quiescent on backs, with occasional movements of legs. Mar. 8.|
|10.0.||Ten flies dead; several others seem unaffected. A large blowfly introduced into the cage, and observed to drink the solution for 15 minutes.|
|10.30.||Blowfly not so active; does not fly away when probed; shows what seem to be involuntary movements of the legs.|
|10.40.||Blowfly very lethargic; walks heavily; can be pushed along without offering to fly; every now and then one or other wing extended to full extent, and held there a moment; this is not a voluntary movement; does not seem to have proper control of legs when walking—one or more legs move irregularly, and not in a suitable direction.|
|11.0.||Blowfly allowed to escape from cage; remained in one position on table till touched, and then flew heavily on to window, crawled, into a corner, and was subsequently lost sight of. Two other blowflies introduced into cage; one immediately fed from the watch-glass, and in 15 minutes showed symptoms; lethargic, and disinclined to move.|
|12.0.||Second blowfly has been observed to drink but sparingly. First blowfly showing more marked symptoms; tonic spasms of wings and uncontrolled movement of legs; allowed to escape, but on attempting to fly seized with a general convulsion, buzzing round and round on back; was attacked with seizures of this kind until exhausted.|
|1.0.||Lying on side, dead. Quite a considerable drop of clear fluid has exuded from proboscis.|
|Mar. 10.||All the house-flies are dead, and all show marked distention of abdomen. Blowfly apparently unaffected, so allowed to escape; flew about on window for few minutes, and was then seized with convulsions and died, as the others had done.|
|Jan. 30.||At 4.30 p.m. some mince was allowed to become infected by blowflies. A small portion of the meat containing the larvæ was placed m each of four watch-glasses, and covered with a 0 01-per-cent., a 0 025-per-cent., and a 0 05-per-cent. solution tutin, and with normal salme respectively. In each case the larvæ began to wander and coil up in the fluid. After one hour these fluids were poured off, and each watch-glass covered with another, to prevent evaporation.|
|" 31.||At 4.45p.m. the watch-glasses were examined. In, the one treated with normal saline only the larvæ are numerous, have grown, and are very active, crawling all over the surfaces of both watch-glasses. In the 0 01-per-cent. and 0.025-per-cent. solution only one or two larvae are, to be seen in movement; they are very small, and have not wandered from the mince. In the 0 05-per-cent. solution no movement is to be seen.|
A drop of hay-infusion, containing paramœcia, amœbae, monads, and, bacteria, was placed on each of four slides, labelled A, B, C, D.
|5.49.||One drop normal saline added to B.|
|5.50.||One drop of a 0.1-per-cent. solution tutin in saline added to C.|
|5.51.||One drop of a 0 1-per-cent. solution quinine hydrobromate in normal saline added to D.|
|65.||B: No change. C: No change. D: No paramœcia to be seen, but monads and bacteria still moving.|
|6.30.||B: No change. C: Paramœcia may still be seen moving, but their movements are irregular; the monads are collecting at the surface and edge of the drop; this may also be seen in A and B Another drop of a 0 1-per-cent. solution added to C. D: No movement.|
|6.50.||A and B: No change. G: Still one or two paramoacia moving; are not so easy to find; many monads still active, others stationary; amœbae still moving; field does not look so lively as it did.|
|7.30.||A drop of hay-infusion placed on a slide, and covered with a cover-glass, and a drop of a 0 1-per-cent. solution tutin normal saline placed at edge of cover.|
|7.35.||Some paramœcia have swum out into drop of tutin solution; at once display irregular movements; remain in the drop of tutin solution.|
|7.40.||Movement of paramœcia in drop of tutin solution, which at first were excited, now becoming slower and very irregular; they roll over and over like rotifers.|
|7.45.||Paramœcia at a standstill in tutin' solution: they appear to disintegrate internally, and are no longer recognisable; paramœcia under the cover-glass quite normal.|
Experiment repeated, using a 0.5-per-cent. solution tutin in distilled water, with similar results.
This experiment was essentially similar to Exp. 119.
|Jan. 28.||Six test-tubes were taken, and labelled—“0.5 per cent, tutin,” “0.4 per cent, tutin,” “0 3 per cent, tutin,” “0 2 per cent, tutin”, “0 1 per cent, tutin,” and “normal saline.” The volume of fluid in each tube was the same.|
|6.0.||A small piece of fresh mmce was added to each tube, and the tubes left open to the air.|
|Jan. 29.||At 5.45 p.m. the tubes were corked, and placed in an incubator at 40° C.|
|" 31.||Tubes opened and examined. The tube containing normal saline was very offensive. None of the other tubes had the offensive smell of putrefaction; the smell was lather like that of stomach-contents.|
|Feb. 1.||Contents of tubes examined under microscope; all show moving bacteria, but the drop taken from the tube containing normal saline was much more crowded, and the bacteria in it showed a greater variety of form and size.|
Four fermentation-tubes were filled—(1) with glucose solution and tutin (0.25 per cent, of the latter); (2) glucose solution, with 0.1 per cent, tutin; (3) glucose solution alone; (4) water. A few drops of a yeast emulsion were added to each.
|5.45.||Tubes placed in incubator.|
|7.10.||Fermentation in (1) and (3).|
|7.45.||Fermentation in (2) as well, but only half as much gas developed as in (1).|
|9.30.||(1), (2), and (3) fully fermented.|
|4.40.||Two fermentation-tubes set on—(1) with glucose alone; (2) with 0 05 per cent. tutin.|
|6.10.||Both equally fermented.|
|8.30.||Both fully and equally fermented.|
Four fermentation-tubes filled with the following:—A: Water (16 c.c.), 25 per cent. glucose (4 c.c.); B: Water (20 c.c.); C: Water (8 c.c.), plus 0.5 per cent, tutin solution (8 c.c.) plus 25 per cent, glucose (4 c.c.) (equals 0.2 per cent, tutin in mixed fluid); D: 0.5 per cent, tutin (16 c.c.), plus glucose (4 c.c.) (equals 0.4 per cent, tutin in mixed fluid). A few drops of freshly procured emulsion of brewers' yeast added to each; D received rather more than the others.
|3.30.||Tubes placed in incubator.|
|4.40.||Small bubbles forming in A, C, and D.|
|6.0.||Fermentation most distinct in D.|
|7.10.||Most fermentation in D; slight in C; little or none in A and B.|
|7.30.||Added another drop of yeast emulsion to A.|
|8.0.||Fermentation now as great in A as in D.|
This shows that the amount of yeast added exerts a greater influence on the amount of fermentation that occurs than does the presence of tutin.
Five test-tubes were taken, containing a 0.1-per-cent., a 0.2-per-cent., a 0.3-per-cent., a 0.4-per-cent., and. a 0.5-per-cent. solution tutin in normal saline; and one test-tube containing normal saline alone. Equal volumes of fluid in each test-tube.
|Jan. 23.||At 4.30 p.m. twelve mustard-seeds were placed to soak in each test-tube.|
|"23.||At 4.30 p.m. the seeds from each test-tube were sown on moist felt placed in separate tin boxes, correctly labelled.|
|"24.||All the seeds in each box have germinated.|
|"25.||At 10.10 a.m. the same progress has been made in each case. All the seeds were replaced in their respective solutions.|
|Jan. 26.||The seeds were resown in the tin boxes.|
|" 30.||All have grown more or less. The seeds soaked in normal saline have made most progress, and are closely followed by those soaked in 0.1 per cent, and 0.2 per cent. Those soaked in 0.4 per cent, and 0.5 per cent, are growing, but only two or three seeds in each, and the growth is not so vigorous as in the others. Since the 26th the felt in the boxes has been kept moist by the addition of water.|
|Feb. 3.||All the seeds soaked in normal saline have grown, and are growing vigorously. In the boxes labelled “0.1 per cent.” and “0.2 per cent.” three or four seeds-are growing vigorously, and have made nearly as much progress as in the box labelled “normal saline,” but the others remain stunted, and one or two have made no progress since being resoaked in the tutin solution.” In the boxes labelled “0.4 per cent.” and “0.5 per cent.” one or two seeds have made good progress, but most of the others are stunted, or have not grown at all.|
A few bars were taken from the gill of a cockle, and mounted in sea-water, and observed under the low power.
|4.0.||Normal saline perfused under cover-glass.|
|4.35.||No appreciable difference.|
|4.38.||0.1-per-cent. tutin solution perfused under cover-glass.|
|4.46.||0.2-per-cent. solution tutin perfused under cover-glass.|
|4.50.||Seems to be a slight retardation, but doubtful.|
|4.51.||0.3-per-cent. tutin solution perfused under cover-glass.|
|4.52.||Obvious slowing; many cilia stationary, others moving feebly.|
|4.59.||Ciliary movement practically ceased.|
|5.10.||Fresh specimen taken in 0.3 per cent.|
|5.13.||Very feeble movement.|
Normal saline and afterwards sea-water were then perfused under cover-glass, but recovery did not take place.
|0.30.||Fresh specimen taken, and a 0.2-per-cent. solution tutin perfused under coverglass.|
|5.37.||Movements have practically ceased, except for a few cilia here and there.|
|5.45.||All movement apparently ceased.|
A solution of 1/1000 KOH in sea-water perfused under cover-glass, and ciliary movement immediately resuscitated.
|5.54.||Fresh specimen taken, and a 0.2-per-cent. solution tutin applied.|
|6.5.||Cilia still moving; movement confined to the apices of the cilia. A drop of 0.3-per-cent. solution tutin applied.|
|6.15.||Still moving, but less actively. A drop of 0.5-per-cent. solution tutin applied.|
As this piece of gill was rather large, a fresh specimen was taken from a mussel.
|7.45.||0.2-per-cent. solution tutin applied.|
|7.52.||A distinct slowing has occurred, but the cilia are still able to cause a movement of particles. This only occurs in sheltered places. The cilia are at a standstill on the edges.|
|7.57.||A drop or two of sea-water applied, and many cilia restored to action. In every case a control mounted in sea-water was used for comparison.|
Two or three gill-bars from a small rock-oyster were taken and mounted in seawater under a cover-glass. Two preparations were made, and one used as a control. (Low power.)
|2.30.||A drop or two of a 0.5-per-cent. solution in sea-water tutin perfused under cover-glass.|
|2.35.||Appears as if there was a slight slowing of cilia in exposed situations, but doubtful.|
|2.40.||Cilia still moving; more 0.5-per-cent. solution tutin perfused under cover-glass.|
|3.0.||Most cilia in active movement; here and there a few are motionless, or moving feebly.|
A fresh preparation was made from the gill of a cockle, and examined with the low power without a cover-glass. A control was used.
|3.10.||A drop of a 0.5-per-cent. solution tutin applied.|
|3.12.||If anything, movements seem exaggerated.|
|3.20.||First drop tutin solution dried off, and a second drop added.|
|3.50.||No effect observable.|
|4.30.||Still in active movement.|
Three preparations of ciliated epithelium from the gullet of a frog. No. 1 was mounted in normal saline without a cover-glass, and used as a control; No. 2 was mounted in a drop of a 0.1-per-cent. solution tutin, without a cover-glass: No. 3 was mounted in a drop of a 0.5-per-cent. solution tutin, without a coverglass. Three microscopes were used, and the specimens examined under the low power, with the draw-tube out.
|5.14.||Moving.||No effect.||No change.|
|6.10.||The specimens had partially dried, and cilia were seen moving in preparation 3 only.|
The gullet of a frog was exposed, and the rate of progress along it (by ciliary action) of a small fragment of cork was observed. A centimeter scale was arranged to lie parallel with the gullet, and the time the cork took to travel 1 cm. noted as follows:—
|Normal||Gullet bathed with a 0.3-per-cent. Solution Tutin.|
|Average, 50.4″||Average, 34.8″|
Two frogs' nerve-muscle preparations made. Muscles placed in watch-glasses with normal saline. Nerve laid on frog-plate, and kept moist with saline, and stimulated with break shocks. A (m saline), minimal stimulus found to be with coil at 17 cm. B (in saline), minimal stimulus at 21.5 cm. B was then placed in a watch-glass cortaming a 0.025-per-cent. solution tutin in normal saline at 5.54 p.m.
|5.55.||Contracts at||20 cm.||20 cm.|
|6.8.||"||20 cm.||18 cm.|
|6.17.||"||21 cm.||19.5 cm.|
|7.15.||"||19.5 cm.||18 cm.|
|7.35.||"||19 cm.||17 cm.|
|8.10.||"||19 cm.||16 cm.|
|8.52.||"||18 cm.||15 cm.|
|9.35.||"||18 cm.||12 cm.|
In this experiment, the nerve of the one preparation (A) was placed in tutin solution (0.5 per cent.), and the muscle of the other (B) in the same. Before placing in tutin solution, A contracts at 28 cm., and B at 34 cm.
|6.3.||Tutin solution applied as above.|
|6.6.||Contracts at||31.5 cm.||28.5 cm.|
|6.17.||"||30.5 cm.||30 cm.|
|7.22.||"||33 cm.||27.5 cm.|
One muscle in saline (B) and one in a 0.05-per-cent, solution tutin (A). Nerves exposed to air, and kept moist. Stimulated as before. Before tutin applied. A contracts at 30.5 cm., and B at 25.5 cm.
|6.15.||Placed in tutin solution.|
|6.20.||Contracts at||28.5 cm.||25 cm.|
|6.28.||"||28.5 cm.||24 cm.|
|7.25.||"||24.6 cm.||22.6 cm.|
|8.20.||"||25.5 cm.||25 cm.|
Same as previous experiment. Before tutin (interrupted current), A contracts at 36 cm., and B at 32 cm.; single shocks, A contracts at 27 cm., and B at 25.5 cm.
|10.56.||A placed in a 0.05-per-cent, solution tutin in normal saline. B left in saline.|
|11.2.||Contracts at||27 cm.||26 cm.|
|11.20.||"||26.5 cm.||25 cm.|
|12.20.||"||24 cm.||24.5 cm.|
|1.0.||"||21.5 cm.||23 cm.|
Rabbit (young). Weight, 804 grams.
Blood-films were taken, dried in the air, and fixed with Jenner's stain; and next morning at—
|11.36½.||Gave 25 minims of a 0.5-per-cent, solution tutin (10 mlgm. per kilo), hypodermically.|
|11.56.||Shaking-movements of head; pupils normal.|
|11.58.||Shaking and rocking of whole body; lying prone on belly.|
|11.59.||Twitching of ears.|
|12.0.||Twitching of hind limbs and then of fore limbs.|
|12.1½.||Severe convulsions; kicking-movements; head drawn back; clawing-movements; convulsions lasted 2 minutes; pupils did not dilate; cornea damaged by rubbing open eye on floor in convulsion.|
|12.3½.||Lying on side, with head bent back; hind and fore limbs show running-movements.|
|12.6.||Convulsion passed, but running-movements of legs continue.|
|12.11.||Convulsion passed; running-movements continue; head bent well back.|
|12.13.||Convulsive movements of jaw and ears, gradually merging into a general convulsion.|
|12.18.||Jaw and limb movements continue. The lower jaw is pushed forwards, and then clenched so that lower incisors lie outside upper incisors, then they slip inside the upper incisors with a grating sound.|
|12.22.||Movements still continue; no cry has been uttered by the animal as yet; respiratory movements are not much in evidence; movement of limbs, jaw, and retraction of head most prominent.|
|12.32.||Same; lying on left side, with occasional kicking-movements.|
|12.31.||All movement ceased, except that heart can be seen beating by movement of hairs over precordial region.|
P.M.—Stomach not distended; contained only food; blood-film taken from the left ventricle only; heart had ceased beating; lungs not congested; bladder full.
Result.—10 mlgm. per kilo caused death m less than one hour.
Experiments 140 to 151 inclusive referred to the blood-pressure work, details of which are omitted.
|P.M.||Cat. Weight, 3.52 kilograms.|
|3.6.||Chloroform administered with a. Skinner's mask. The skin was reflected, the cranium trephined over the right parietal region, and the bone removed with bone forceps until the greater part of the right cerebral hemisphere was exposed.|
|4.10.||50 minims of a 0.5-per-cent, solution tutin was injected hypodermically, and the administration of the anæsthetic discontinued.|
|4.24.||As no symptoms had appeared, 25 minims of a 0.5-per-cent. solution tutin was injected into the peritoneum.|
|4.32.||Slight twitching of the head and ears noticed, followed in a minute or two by convulsive movements of the paws. The right cerebral hemisphere was immediately scooped out. The convulsive movements continued as before, affecting both sides equally. The left cerebral hemisphere was then removed. The convulsive movements continued. The breathing was greatly exaggerated, and at intervals large quantities of urine were shot out with considerable force.|
|4.45.||The spinal cord was then divided at the level of the 5th dorsal vertebra.|
|4.49.||Spasmodic movements of the hind limbs; the limbs were rigidly flexed on the abdomen and shaken with clonic spasms.|
|4.54.||Movements of fore paws.|
|4.56.||Slow spasmodic movements of hind limbs, the slow rigid movement towards the abdomen ending in clonic spasms. The reflexes in the hind limbs are exaggerated, the legs being quickly drawn up, and showing clonic movements.|
|5.0.||The fore limbs have been quiescent for some time; movements occurring in the hind limbs only.|
|5.3.||Reflexes still markedly exaggerated (clonus) in the hind limbs, but cannot be elicited from the fore limbs.|
|5.9.||All respiratory movements ceased, and reflexes absent.|
P.M.—The chest was opened at once, and slight heart-movements found to be still present. The bladder, notwithstanding the large quantities of urine ejected during the experiment, was full. The cerebral hemispheres were found to be entirely removed; the corpora quadrigemina were intact; the cord was found to be divided within the dura mater at the level of the 5th dorsal vertebra; the cord was crushed through rather than cut through (scissors, not too sharp, having been used to divide it), but its continuity was completely destroyed.
|P.M.||Cat. Weight, 2.4 kilograms.|
|2.50.||Chloroform ana [ unclear: ] esthesia induced, and spinal cord exposed, and completely divided in the mid-dorsal region; 1 c.c. of a 0.5-per-cent solution of tutin was then injected into the peritoneum, and chloroform discontinued.|
|3.0.||No response to electric stimulus applied to left hind foot, but marked response when applied to left fore foot.|
|3.5.||Slight twitching of ears and jerking of head.|
|3.14.||Distinct tutin twitches of head and fore limbs.|
|3.16.||Convulsion of fore part of body, accompanied by movement of tail; hairs of tail have not become erect; great salivation.|
|3.20.||Convulsive movement of fore part of body continues; a movement of the tail near the tip; 1 c.c. of a 0.5-per-cent. tutin injected into peritoneum.|
|3.22.||Swishing-movement of tail from side to side; reflex response to pricking left hind leg with a knife distinctly obtained during convulsion of fore part.|
|3.28.||Movement of tail during convulsion of fore part.|
|3.29.||Severe convulsions of fore part, accompanied by marked lashing-movement of tail.|
|3.32.||More convulsions, with lashing of tail; movement of tail sometimes occurs quite apart from any movement in fore part of body, but occurs also with marked regularity at the beginning of each fit of fore part.|
|3.42.||Several movements of tail, with no convulsion of fore part; tail sometimes moved from root and sometimes only the tip; movement is mainly from side to side, but now and then the whole tail is lifted upwards. Reflexes present in the hind part; movements of both limbs on tapping one.|
|3.45.||Movement of tail becoming more marked, while fore part more quiescent.|
|3.47.||Hind limbs spasmodically drawn up to abdomen several times; then a slight fit of fore part occurred, and movements of tail, with tetanic movement of hind limbs.|
|3.50.||Tonic and clonic spasm of hind limbs; claws extruded.|
|3.52.||Respirations slow and gasping.|
|3.54.||Heart-beat vigorous; as heard by stethoscope, 72 per minute. Twitches of fore paws and tonic spasm of hind limbs, with erection of hairs of tail.|
|4.4.||Clonic movements of left hind limb.|
|4.5.||Distinct tonic and clonic spasm of both hind limbs; limbs fully extended, with claws extruded; respiration infrequent and difficult, owing, to obstruction of air-passages by saliva and mucus; heart intermittent, misses one or two beats towards the end of inspiration.|
|4.6.||More movements of hind limbs, fore part motionless; respirations about four per minute.|
|4.6½.||Another spasm of hind quarters.|
|4.7.||Respirations ceased; heart audible by stethoscope, beating regularly for two minutes and a half after last respiratory gasp.|
A frog (Hyla aurea), pithed in the ordinary way three hours previously, was suspended by the head. At intervals of a few minutes its, feet (as far as the ankle) were dipped into dilute sulphuric acid, of a strength of 1 in 1,000. After each dipping the feet were carefully washed in fresh water. The time was taken by a seconds-clock, and the result was as follows:—
|Right Foot.||Left Foot|
6.3 minims of a 0.1-per-cent, solution tutin(11 mlgm. per kilo) was then injected, under the skin of the back, and after an interval of 30 minutes the feet were again dipped in the acid:—
|Right Foot.||Lett Foot.||Frog A|
Here a further dose of 10 minims was injected, and after an interval of 15 minutes the feet were dipped as before:—
|Right Foot.||Left Foot.||Forg A.|
Each, time the feet were withdrawn together, and, it was thought, more actively than before the tutin was injected.
The experiment was repeated with a second frog, pithed in the same way four hours previously. The result was as follows:—
|Right Foot.||Left Foot.|
10 minims of a 0.1-per-cent. solution tutin was then injected under the skin of the back, and after an interval of half an hour the reaction-time again tested. The time when movement first took place only was noted.
|Twitched at 2″;||withdrawn at 4″|
At 22 seconds there was a slight twitch, but no further movement took place, although 70 seconds were counted. Pinching or pricking a foot met with an immediate response, but the acid had no influence. Five minutes later no reflex movement could be elicited by any means, and the heart could no longer be seen beating through the chest-wall.
Frog A was then tested again, and found to respond as actively as before.
A third frog, pithed one hour previously, was treated in the same way.
|Frog C||10″||Right leg only withdrawn. Frog removed from acid as soon as first movement took place.|
2 minims of a 0.5-per-cent. solution was then injected under the skin of the back, and 10 minutes were allowed to elapse before the feet were dipped again.
|Frog C||6″||Both legs withdrawn.|
1 minim of a 0.5-per-cent. solution tutin was then injected into the heart.
|A fourth frog, pithed one hour previously.|
|Frog D||8″||Movements of lower limbs only.|
5 minims of a 0.1-per-cent. solution tutin (10 mlgm. per kilo) was then injected under the skin of the back, and six minutes later the test reapplied.
|Frog D||2″||Very extensive general movements.|
|"||4″||Arms moved as well.|
|"||2″||Legs and subsequent movements of arms.|
|"||1″||Legs, spreading to arms.|
|"||2″||Legs, arms not moved so vigorously.|
|"||4″||Both legs only.|
|"||5″||Legs only, response more sluggish.|
|"||2″||Legs and arms.|
The experiment was repeated a fifth time. In this case the frog had been pithed in the usual way one hour previously, but it appeared to retain some power of voluntary movement, and was so restless that it was suspected that the cerebrum had not been completely destroyed. Extensive general movement followed each application of the acid.
|Left Foot||Right Foot.|
1 minim of a 0.5 per-cent. solution tutin was then injected (about 12 mlgm. per kilo) under the skin of the back, and 15 minutes later the feet were again dipped into the acid; but before this was done a typical tutin convulsion occurred.
|Left Foot.||Right Foot.|
|"||50″||70 seconds were counted, but the right foot was not withdrawn.|
On examination the optic lobes were found to be intact.
The experiment was repeated with a sixth frog. This was beheaded a little in front of the anterior border of the tympanic membrane, so as to sever all in front of the top of the medulla.
|Right Foot||Left Foot|
1 minim of a 0.5-per-cent. solution was then injected under the skin of the back, and 8 minutes later the test reapplied.
|Right Foot||Left Foot.|
As a result of letting the frog fall on the table, a typical tutin spasm, with croaking, occurred here. Seemed to require severe irritation to induce a spasm. A spasm occurred on striking sharply with a glass rod.
250 seconds were counted, but feet not withdrawn. Slight twitches occurred, however, at 58″, 71″, 86″, 94″, and 120″. 100 seconds more, but no effect.
Although the acid had ceased to have any influence, it was still possible to elicit reflex action on painful stimulation—e.g., pinching.
Ten minutes later reflexes could not be elicited in any way. The thorax was then opened, and the heart found to be still beating. The auricles were distended. Twitching occurred as the cord was being pithed.
A few drops of a solution of tutin were introduced into the right eye of a young rabbit. Within a few minutes there seemed to be a very slight dilatation. In an hour and a quarter the right pupil was thought to be slightly the larger by an observer ignorant of which eye had been subjected to the test, but the difference was so very slight, if existing at all, that the result was regarded as doubtful.
Four small drops of a 0.5-per-cent. solution tutin were introduced into the left conjunctival sac of a rabbit. It was observed for 15 minutes, and no change in the size of the pupil was noted. No hyperæmia of the conjunctiva resulted, nor did the conjunctiva become less sensitive.
This rabbit died twenty-four hours later, after exhibiting symptoms of tutin poisoning.
|2.50.||Two drops of a 0.5-per-cent. solution of tutin introduced into the right eye of a cat.|
|2.52.||No apparent effect.|
|2 55.||No change.|
|3.5.||Pupils are the same size, and react equally to light.|
|3.25.||0.5 c.c. of a 0.5-per-cent. solution of coriamyrtin was instilled into the right eye of a rabbit.|
|3.30.||No apparent effect.|
|3.35.||Difficult to say if any effect; the right eye may possibly be slightly contracted.|
|3.40.||An observer ignorant of which eye had been treated was unable to distinguish any difference between the two pupils.|
|4.0.||Pupils appear equal, and react to light with equal readiness. No appearance of irritation of conjunctiva.|
|3.14.||Two drops of a 0.5-per-cent. solution of coriamyrtin introduced into the left eye of a cat.|
|3.16.||No change in size of pupil observed.|
|3.30.||Pupils equal, and react to light with equal readiness.|
|4.0.||Pupils appear to be equal.|
Rabbit. Weight, 777 grams.
|Jan. 24.||At 4.50 gave 13.2 minims of a 0.1-per-cent. solution tutin (1 mlgm. per kilo), hypodermically.|
|" 25.||No obvious effect.|
|" 27.||Gave 13.2 minims of a 0.1-per-cent. solution, per os.|
|" 28.||Normal, so gave 13.2 minims of a 0.1-per-cent. solution, per os.|
|" 30.||Repeated dose of 13.2 minims of a 0.1-per-cent. solution, given per os.|
|Feb. 4.||Weight, 848 grams. Gave 21.6 minims of a 0.1-per-cent. solution (1.5 mlgm. per kilo), per os.|
|" 8.||Normal. Gave 4 minims of a 0.5-per-cent. solution tutin (1.5 mlgm. per kilo), per os.|
|" 11.||Weight, 879 grams. Gave 2 mlgm. per kilo, per os.|
|" 12.||No symptoms; normal.|
|" 14.||Weight, 862 grams. Gave 7.3 minims of a 0.5-per-cent. solution tutin (2.5 mlgm. per kilo), per os.|
|" 17.||Weight, 922 grams. Normal, so gave 7.8 minims of a 0.5-per-cent. solution tutin (2.5 mlgm. per kilo), per os.|
|" 24.||Weight, 958 grams. Normal, so gave 9.7 minims of a 0.5-per-cent. solution (3 mlgm. per kilo), per os.|
|" 27.||Weight, 983 grams. Gave 11.6 minims of a 0.5-per-cent. solution (3 mlgm. per kilo), per os.|
|Mar. 3.||Weight, 991 grams. Gave 13.4 minims of a 0.5-per-cent. solution (4 mlgm. per kilo), per os.|
|" 7.||Normal. Gave 5 mlgm. per kilo, per os.|
|" 12.||Weight, 1,154 grams. At 2.40 p.m. gave 23.5 minims of a 0.5-per-cent. solution tutin (6 mlgm. per kilo), per os. At 4 p.m. shows symptoms; extends fore and hind limbs, and lies on belly; ears and head tremulous.|
|" 16.||Weight, 1,181 grams. Gave 7 mlgm. per kilo. Not continuously observed. Recovered.|
|" 19.||Weight, 1,176 grams. Gave 8 mlgm. per kilo. In 3 ½ hours showed symptoms (twitching of ears, chewing-movements, &c.). Recovered.|
|" 24.||Gave 11.6 mlgm. per kilo, per os. Result, death in two hours.|
Guinea-pig. Weight, 627 grams.
|5.0.||Gave 10.2 minims of a 0.1-per-cent. solution of tutin (1 mlgm. per kilo), hypodermically, with antiseptic precautions.|
|6.30.||Convulsive movements of jaw; animal standing in unnatural attitude.|
|Jan. 25.||Apparently normal.|
|" 27.||Gave 10.2 minims of a 0.1-per-cent. solution tutin (1 mlgm. per kilo), per os.|
|" 28.||Showed no symptoms; normal to-day, so repeated dose, 10.2 minims per os.|
|" 30.||No symptoms; repeated dose, 10.2 minims, per os.|
|Feb. 4.||Weight, 644 grams. At 5.36 p.m. intended to give 17 minims of a 0.1-per-cent. solution tutin (1.5 mlgm. per kilo), per os, but syringe not set before giving, and so probably less than 17 minims given.|
|5.50.||Intended to give 17 minims of a 0.1-per-cent. solution tutin, per os, but 17 minims of a 0.5-per-cent. solution was given by mistake. Mistake discovered, and 0.5-per-cent. gram chloral-hydrate given, per rectum, at 6.5 p.m. Immediately on withdrawing nozzle of syringe a few pellets of fæces were discharged, but very little fluid returned.|
|6.20.||Is under the influence of chloral, but showing twitches of fore limbs.|
|7.5.||Apparently still under chloral; is quite limp.|
|9.0.||Awake, and easily startled; ears cold, and whole body shivering; was wrapped in towel.|
|9.20.||Will not remain covered up. Shivering not so marked.|
|9.0.||Apparently normal, quite warm, breathing is regular.|
|2.0.||Does not seem to have eaten; seems dazed.|
|11.0.||Did not try to escape when went to lift it, but resents being handled; otherwise appears normal.|
|9.0.||Seems stiff in the hind quarters; moans when touched; has not eaten. Noticed moist condition of uroanal opening.|
|2.30.||Completely paralysed, and cold in hinder half of body; breathing thoracic; eyes kept wide open; moans when handled, and showed some convulsive movements of fore limbs and head.|
P.M.—Stomach distended with gas; intestines contain mixed gas and fluid contents; peritoneum much injected; appearance suggestive of general peritonitis.
Cat. Weight, 2.686 kilograms.
|5.12.||Gave 13 minims of a 0.13-per-cent. solution of picrotoxin (0.375 mlgm. per kilo), hypodermically.|
|5.22.||Quite normal; cleaning itself.|
|5.40.||Same. No departure from the normal.|
|Feb. 28.||Quite normal.|
|Mar. 3.||Same cat. Weight, 2.699 grams. Fresh solution of picrotoxin|
|4.0.||Gave 6.8 minims of a 0.25-per-cent. solution picrotoxin, freshly prepared (0.375 mlgm. per kilo), hypodermically.|
|4.20.||Has defæcated, and swallows occasionally, but seems so well that these may be normal appearances.|
|5.15.||No apparent abnormality.|
|Mar. 4.||Has displayed no symptoms, and seems perfectly well.|
Cat. Weight, 2.699 kilograms.
This cat previously received 1 mlgm. tutin, and recovered; will now receive an equimolecular solution of picrotoxin, 887/336 × 1 mlgm. picrotoxin = 2.66 mlgm. picrotoxin = 18 minims of a 0.25-per-cent. solution picrotoxin.
|5.20.||Gave 18 minims of a 0.25-per-cent. solution of picrotoxin.|
|6.0.||No symptoms as yet, beyond occasional swallowing.|
|6.7.||Salivating; mouth open, panting, vomited; vomiting repeated several times; breathing slower after vomiting; defæcated|
|6.10.||Retching; this a more marked symptom than with tutin. Animal looks much less disturbed than with tutin at this stage.|
|6.15.||Every now and then spasmodic contraction of diaphragm, suggesting hiccough; looks sleepy.|
|6.20.||Twitching affecting fore paws only; no face or ear twitching.|
|6.25.||Twitching again, more marked, causing a little spring with fore part of body; is very easily startled.|
|6.27.||Another more marked twitch; sudden spring, with fore arms bowed, giving appearance of trying to cling to the ground; cat more alarmed and astonished at the twitching than in the case of tutin; cerebrum seems less affected; twitches at the slightest noise—e.g., tap of foot on floor, or turning page of note-book.|
|6.32.||Much more marked twitching; cat stands in unnatural attitude, afraid to move; every movement causes a sharp spasmodic contraction of muscles; breathing very rapid.|
|6.35.||General clonic convulsion, lasting a minute and a half; no tonic stage noticed; rose immediately seizure had passed; stands in unnatural attitude, afraid to move; twitching of eyelids noticed for first time; seems quite clear mentally.|
|6.40.||Very cautiously assumed a sitting-position, on haunches; continuous twitching; looks up at once if spoken to; twitching has not been so general since the convulsion, nor is it so easily excited by stamping the foot; not so much salivation as with tutin; watching with interest movements of a rabbit in a cage opposite.|
|6.50.||Twitching only very occasional and slight; sitting with eyes closed, as if dozing; breathing normal.|
|6.55.||Twitching more marked again, and is again easily induced by a sudden noise.|
|7.12.||Sitting on haunches, in normal attitude; gives occasional uneasy starts; head drawn back quickly, or to one or other side.|
|7.22.||Still twitching a little; eyes half-closed most of the time; no sign of pain.|
|7.33.||Came to front of cage, and rubbed itself against wires; then sat down as before; is still disturbed by twitchings of fore limbs and head, but it is slighter than it was.|
|7.43.||Drowsy, but wakes up with a start.|
|8.0.||Starts are much less frequent, and very slight.|
|8.22.||Apparently asleep, sitting on haunches; no further twitching noticed.|
|Next day. Quite recovered.|
Cat. Weight, 3 kilograms.
|4.38.||Received 10 minims of an emulsion of coriamyrtin (3 mlgm. or 1 mlgm. per kilo).|
|4.48.||Twitching first noticed; saliva dripping from mouth; mouth open.|
|4.50.||Vomited; breathing greatly exaggerated.|
|4.54.||Breathing still rapid; twitching increases.|
|4.56.||General convulsion, mostly clonic, lasted 30 seconds; pupils dilate with each twitching.|
|5.0||Panting hard and twitching constantly.|
|5.1.||General tonic and clonic convulsion, emprosthotonos, almost standing on head; breathing suddenly became normal after convulsion; then lay with face pressed on floor, breathing heavily; minor twitching continues.|
|5.7.||Clonic convulsions, chin bent down on chest, limbs spread-eagle on either side; stands in an elongated crouching attitude, pressing forehead against floor; respirations, 160 per minute.|
|5.8.||Clonic convulsions, emprosthotonos, pupils return to normal size in intervals.|
|5.10.||Another convulsion; pupils dilated, as usual.|
|5.14.||General convulsions; intervals very short.|
|5.20.||During a fit, tail rigidly arched over back; died in this convulsion. Heart inaudible to stethoscope immediately after.|
P.M.—Three hours afterwards. Membranes of cord and brain markedly congested; intestines pale; heart, right side enlarged, left side contracted; hæmorrhagic patches in lung; urinary bladder empty; gall-bladder moderately full; uterus contained three fœtuses; pieces of cord and brain fixed in 8 per cent. formal sections, made by paraffin method.
Rabbit. Weight, 1.264 kilograms.
|4.47.||Gave 0.6 grams of a 1.in-1 solution chloral-hydrate, per rectum. Slight loss of the fluid.|
|4.57||Is anæsthetised; gave 17.1 minims of a 0.5-per-cent. solution of tutin (4 mlgm. per kilo), hypodermically.|
|6.25.||Recovered consciousness; marked twitching of head and ears; sitting up.|
|7.15.||Sitting erect on fore paws; salivating freely; convulsive movements of jaws.|
|7.25||Seizures almost continuous; placed on a table; it raises itself by straightening to its fore paws, and pushes its body further and further back, then sinks forward|
|7.45.||apparently exhausted, so that the fore limbs are spread out at right angles to the trunk; meanwhile jaws are grinding together, and saliva seems to be swallowed; pupils moderately dilated, but as daylight nearly gone this may be natural.|
|7.57.||As condition continues, gave another rectal dose of 0.6 gram chloral. After getting this it showed the symptoms described by Marshall, of pushing itself back till it actually fell backwards over its tail. The pupil was dilated, and would not contract when a match was held close to it.|
|8.7.||No cessation of symptoms; fell over tail again; interval between seizures varies, about one minute; no general convulsion, as is the case when chloral is not given.|
|8.15.||Has been quieter the last few minutes, as if dozing as it sits.|
|8.30.||Lying on side, apparently under chloral; breathing very shallow; no twitching.|
|8.40.||Respirations extremely feeble.|
|8.45.||Spontaneous movements; tried to rise; raised head, and fell over on the other side.|
|9.0.||No further tutin symptoms; moves occasionally, especially if touched.|
|9.30.||Still the same; has passed urine; is sitting with nose in a corner of cage, as if dozing; eyes half-shut; does not mind being touched or handled; ears cool.|
|10.0.||Same, but has changed its position several times.|
|10.40.||Moving about more freely; seems to suffer from irritation of the anus; is more like normal than it has been since 7 p.m.|
|10.50.||Same; has passed some pellets of fæces and some thin fæcal matter; behaves normally when approached, and seems wide awake; can use all its limbs.|
|Feb. 12.||At 5 p.m. seems normal, except that it is suffering from diarrhœa.|
|" 13.||Found dead in the morning.|
Rabbit. Weight, 1.455 kilograms.
|5.45.||Gave, per rectum, 0.6 gram chloral-hydrate.|
|5.55.||Gave 13.8 minims of a 0.5-per-cent. solution tutin (3 mlgm. per kilo), hypodermically.|
|6.10.||Animal apparently fully anæsthetised; wrapped in a soft towel to maintain temperature.|
|7.5.||Dazed and sleepy looking; no movements other than respiratory; eyes open.|
|7.35.||Made starting-movements with head, and moved hind limbs; is still wrapped in towel; removed towel.|
|8.0.||Has had frequent startings, but is very quiet most of the time.|
|8.25.||Sitting up, apparently dozing; has been somewhat restless, but no sign of convulsion; covered it up with towel, and noticed that it has passed urine.|
|8.55.||Sitting in a corner of cage, quite awake, but does not seem at ease; still frequent startings; breathing slow and deliberate.|
|9.7.||Seems to feel some irritation at anus, possibly from the rectal injection; put its head down, as if to lick anus.|
|10.0.||Has been continuously observed, but no sign of convulsions.|
|11.0.||Apparently quite normal; resists having its ears pulled; has passed some fæces.|
|11.30.||No further change; sitting with eyes wide open; easily startled.|
|Next day. Apparently normal.|
Rabbit. Weight, 1.534 kilograms.
|3.15.||Gave 1 c.c. (1 gram) of a solution of chloral-hydrate, per rectum, and 20.8 minims. of a 0.5-per-cent. solution tutin, hypodermically (4 mlgm. per kilo).|
|3.25.||Lying on side, under chloral anæsthesia; respiration, 42 per minute.|
|3.45.||Still sleepy under chloral; breathing the same; no tutin symptoms.|
|3.49.||Turned over on to the other side; respirations, 40 per minute; blinks eyes once or twice in succession occasionally.|
|5.0.||Twitching and convulsive movements.|
|5.15.||Same convulsive movements.|
|6.30.||Same, but more feeble.|
Exps. 176, 177, 178, 179.
Four frogs. Weights: A, 27 grams (Exp. 176); B, 37-5 grams (Exp. 177); C, 28 grams (Exp. 178); D, 34.5 grams (Exp. 179).
6.0. To each frog was given 0.012 gram chloral-hydrate, hypodermically. It was intended to give 0.008 gram chloral-hydrate to frogs C and D, but by mistake they received the same dose (0.012 gram) as A and B.
6.5. B received 15.3 minims of a 0.05-per-cent. solution tutin (12 mlgm. per kilo), hypodermically; C received 10.4 minims of a 0.05-per-cent. solution tutin (11 mlgm. per kilo), hypodermically; D received 14 minims of a 0.05-per-cent. solution tutin (12 mlgm. per kilo), hypodermically.
Next day. A is normal, B is dying, C is dead, D is dying.
P.M. Rabbit. Weight, 1.4 kilograms.
5.55. Gave 1.5 grams urethane, by stomach-tube.
6.20. Apparently under, very sleepy and limp. Gave 4 mlgm. tutin per kilo, hypodermically.
7.30. Symptoms first observed; they were not present at 7.15 p.m.
8.0. Twitching of ears and head, and starts of fore part of body; no salivation; was handled a little roughly in trying to see if salivation present, and went into a typical fit; it lay on one side, and fit succeeded fit in rapid succession; movements chiefly clonic; no tonic phase could be detected.
8.30. As fits still continue, and animal seemed doomed, tried about 1 c. c. of a 10-percent. sodium-carbonate, per rectum. While giving injection noticed involuntary urination.
8.45. Sodium-carbonate had produced no obvious effect, so tried chloroform; administered it as carefully as possible, with a Skinner's mask; breathing was shallow and rapid; under influence of chloroform animal became quiet, movements of limbs ceased, head moved slowly backwards, and breathing ceased suddenly without the least warning; the heart could not be felt beating, and artificial respiration had no effect.
Feb. 4. Rabbit was chloroformed, stomach-tube passed; 0.85 c.c. paraldehyde (1.2 c.c. P.M. per kilo), dissolved in 10 c.c. water, given.
5.0. Noticed that reflexes had returned.
Feb. 5. Recovered.
P.M. Rabbit. Weight, 1.551 kilograms.
4.48. Gave 1/200 grain hypodermic tabloid of hyoscine hydrobrom, hypodermically.
5.15. Apparently no effect, so gave a second dose of 1/200 grain, per os.
6.5. No effect.
Next day. Normal.
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