O^l-C^- ^5^5^33"^ Issued September 25, 1912. A 0 ucso U 1 JTHE 0 8 — '=^-^ — S 3I0NAL 4 3 ^^1 2 1 7 U. S. DEPARTMENT OF AGRICULTURE, BUREAU OF ANIMAL INDUSTRY.— Bulletin 153. A. D. MELVIN, Chief of Bureau. HE ACTION OF ANTHELMINTICS ON PARASITES LOCATED OUTSIDE OF THE ALIMENTARY CANAL. BY BRAYTON HOWARD RANSOM, Chief of the Zoological Division, AND MAURICE C. HALL, Assistant Zoologist,- Zoological Division. WASHINGTON: GOVERNMENT PRINTING OFFICE. 1912. Digitized by tine Internet Arciiive in 2Q07 witii funding from IVIicrosoft Corporation littp://www.arcliive.org/details/actionofantli.elnniOOransiala Issaed September 25, 1912. U. S. DEPARTMENT OF AGRICULTURE, BUREAU OF ANIMAL INDUSTRY.— Bulletin 153. A. D. MELVIN, Chief of Bureau. THE ACTION OF ANTHELMINTICS ON PARASITES LOCATED OUTSIDE OF THE ■ ALIMENTARY CANAL. BY BRAYTON HOWARD RANSOM, Chief of the Zoological Division^ AND MAURICE C. HALL, Assistant Zoologist, Zoological Division. WASHINGTON: GOVERNMENT PRINTING OFnCE. 1912. THE BUREAU OF ANIMAL INDUSTRY. Chief: A. D. Melvin. Assistant Chief: A. M. FxIRBington. Chief Clerk: Charles C. Carroll. Animal Husbandry Division: George M. Rommel, chief. Biochemic Division: M. Dorset, chief. Dairy Division: B. H. Rawl, chief. Field Inspection Division: R. A. Ramsay, chief. Meat Inspection Division: Rice P. Steddom, chief. Pathological Division: John R. Mohleb, chief. Quarantine Division: Richard W. Hickman, chief. Zoological Division: B. H. Ransom, chief. Experiment Station: E. C. Schroedeb, superintendent. Editor: James M. Pickens. ZOOLOGICAL DIVISION. Chief: B. H. Ransom. Assistant Zoologists: Albert Hassall, Habby W. Gbaybill, Maubice C. Hall, and Howabd Cbawley. Junior Zoologist: Winthbop D. Fosteb. 2 LETTER OF TRANSMITTAL. U. S. Department of Agriculture, Bureau of Aj^imal Industry, Washington, D. C, May 2, 1912. Sir : I have the honor to transmit herewith, and to recommend for publication as a bulletin of this bureau, a manuscript entitled " The Action of Anthelmintics on Parasites Located Outside of the Ali- mentary Canal," by Dr. B. H. Ransom, Chief of the Zoological Divi- sion, and Mr. Maurice C. Hall, assistant zoologist in the Zoological Division of this bureau. This paper details the results of some attempted medicinal treat- ment of sheep for tapeworm disease, and also summarizes our knowl- edge of the use of anthelmintics against parasites located outside of the intestinal lumen. While the experimental results were nega- tive, the evidence brought together here indicates the need of further work, and this paper is intended to simplify additional work by furnishing a systematic summary of all previous work. It also lias immediate practical utility as an aid in judging the claims made for proprietary medicines. Very respectfully, A. D. Melvix, Chief of Bureau. Hon. James Wilson, Secretary of Agriculture. 3 CONTENTS. Page. Introductory 5 Historical review 6 Somatic and extraintestinaJ tseniasis 6 Distomatiasis 10 Autliors' experiments 16 General summary and ci'iticism 19 Bibliography 20 THE ACTION OF ANTHELMINTICS ON PARASITES LOCATED OUTSIDE OF THE ALIMENTARY CANAL. INTRODUCTORY. As commonly employed, the remedies known as anthelmintics are usually efficacious preparations. Their action is based on the prem- ises that they are poisons which can be taken into the digestive tract of such animals as man, the horse, dog, or cat, in quantities not large enough to poison the host but sufficient to stupefj'^ or kill verminous parasites with which they come in contact. The parasites are usually in the small intestine (intestinal helminthiasis) and occasionally in the stomach (gastric helminthiasis). In order to avoid dilution of the medicine, and also to give a more effective contact with the para- site, the patient is prepared in advance by fasting to empty the stomach and intestine. Finally a purgative is administered to carry out the dead or stupefied worms. All this is comparatively simple And in practice effective. Though anthelmintics may be used very successfully in treatment for parasites in the stomach and small intestine of certain animals, they are, as a rule, less satisfactory when applied to ruminants. Medi- cines administered to ruminants must first pass the first, second, and third stomachs, some or all of which are usually well filled with food and difficult to empty in any reasonable period of fasting, before reaching the usual location of gastric parasites, the fourth stomach, through which in turn the medicines must pass before they reach the small intestine. In some cases, however, gastric and intestinal helminthiasis in ruminants may be treated successfully. Perroncito (1885)^ and Stiles (1902), for example, have reported satisfactory results from the use of various remedies for stomach worms of sheep. Certain experiments have indicated that under the right conditions remedies may pass directly through the first stomach and thus arrive more or less promptly in the second and following stomachs and the intestine. Powers (1909), however, has questioned this, and con- siders medicinal treatment for stomach worms in ruminants unsat- isfactory. As to the treatment of parasites located in the large intestine, it has been found in actual practice, even in animals with simple * References to literature will be found In bibliography at end of bulletin. 5 6 ACTION OF ANTHELMINTICS ON PARASITES. stomachs and short alimentary tracts, such as the dog, that the results from the administration of remedies per orem are generally very unsatisfactory. Stiles and Pfender (1902), for instance, found that thymol, a classic remedy for hookworms, was without effect on whip- worms of the cecum of the dog. On the other hand, Miller (1904) claims to have succeeded in removing whipworms from the dog by the use of oleoresin of male fern. Parasites located in the large intestine are, however, difficult to reach with ordinary anthelmintics administered by mouth. In view of the difficulty of reaching parasites located in the ali- mentary tract beyond the stomach or small intestine, or even in these organs in ruminants, by means of anthelmintics administered through the mouth, it would seem on first thought that treatment with such remedies for worms in the liver, pancreas, muscles, brain, blood, or in other locations outside the direct course of the digestive tract, would be certain to result unsuccessfully. Nevertheless several more or less commonly used anthelmintics have received favorable consideration' in the treatment of verminous parasites located outside the lumen of the stomach and intestine, and the results of the tests made in some cases apparently afford a basis for a belief in their efficacy. HISTORICAL REVIEW. A consideration of the work done on this subject shows that writers, in general, have recognized two lines of work: First, the treatment of somatic and extraintestinal tseniasis where parasites are located in the brain, muscles, liver, or subcutaneous tissue; and second, the treatment of certain forms of distomatiasis, where the parasites are located in the liver or blood. Parasites in the liver are, of course, relatively nearer the usual site of operation of anthel- mintics. To the first line of work we wish to add some notes on the treatment of tseniasis where the parasites are located in the liver and in the pancreatic and bile ducts. In this case the parasite, Thysano- soma actinioides, is also an intestinal parasite, and we use, therefore, the terms extraintestinal and intestinal thysanosomiasis to indicate the two forms of infestation with the adult worm. Since writers along either of the two lines mentioned have usually confined their abstracts and criticism of related work to work along the one line, this review will summarize the two lines of work separately. SOMATIC AND EXTRAINTESTINAL TiENIASIS. Ziirn (1882a) states that after trying for 24 years to find some medication that would cure gid in sheep, he has concluded that suc- cessful treatment of the sort is impossible, and cautions against the administration of poisons, which, in his opinion, results in nothing HISTORICAL REVIEW. 7 but a waste of time and money. It is unfortunate that he did not give his experiments in detail, or at least name the remedies he tried; but from the fact that he speaks of them as poisons it seems reasonable to suppose that he tested the various common anthelmin- tics, all of which are poisons. Curtice (1889a and 1890c) records tests of various remedies in intestinal and extraintestinal thysanosomiasis of sheep, and says: Various tseniafuges were tried with little success. The powdered prepara- tions of pumpliin seed, pomegi"anate-root bark, lioosoo, l^amala, male fern, and worm seed proved of no avail. * * * The presence of taeniae in the biliary ducts is another reason why tseniafuges can not be entirely successful in treat- ment of sheep with T. fimbriata [Th. actinioides]. Any medicine which would affect the taeniae in these ducts would also affect the sheep seriously. It is doubtful whether they can be killed or driven from the ducts. Feletti (1894c) administered ethereal extract of male fern in three cases of .human cysticercosis, as follows: 1. Patient had had Taenia solium. Present symptoms: Convulsions, cepha- lalgia, vertigo, vomiting. Small subcutaneous nodules appeared and grew to the size of olives ; one of these was excised and found to be Cysticercus cellu- losae. No cestodc proglottids or eggs were found on fecal examination. Feletti administered 1 to 3 grams of extract of male fern per diem to the patient until he had given 18 grams. The patient could not stand more. The cysticercl diminished to the size of a wheat grain, but the cerebral trouble did not Im- prove, and the man died a month later. No autopsy was permitted. 2. Patient had 34 subcutaneous nodules. One of these was excised and found to be Cysticercus ccllulosce. No cestode proglottids or eggs found on fecal examination. Feletti at first administered 1 to 1.5 grams of extract of male fern per diem, but this was not supported and the dose was accordingly cut down to 40 centigrams. The total amount administered was 26.5 grams. The nodules diminished until they could not be felt. 3. Patient had had a tapeworm three years before and had not recovered the head in several attempts at removal. Present symptoms : Convulsions, vertigo, and vomiting. For this he was treated with sodium and potassium bromld. Seven nodules developed. One of these was excised and found to be a cysticercus. Feletti administered 60 centigrams of extract of male fern per diem, and In a month the nodules had disappeared. The nervous condition was Improved, but the trouble subsequently recurred and was treated with bromids with good results. Feletti thinks the recurrence was due to brain alterations, but considers that the cysticercl were dead. According to Feletti, these cases demonstrate that extract of male fern kills subcutaneous and muscular cysticercl and that it probably acts upon cerebral cysticercl also. He advises a dose of 30 to 40 centigrams per diem for 30 or 40 days. De Renzi (1908) renewed the interest in this line of work by his report of the administration of extract of male fern in four cases of human somatic taeniasis. "We quote the following abstract of De Renzi's cases from Hall (1909) : 1. The patient had occasion to eat badly cooked pork and uncooked sausage; had an infection with Twnia solium five years previous; had attacks of con- 8 ACTION OF ANTHELMINTICS ON PAKASITES. vulsions and insensibility three years later, and on treatment with a vermifuge had passed a tapeworm with the head. Two months later the patient had stronger convulsions,* dizziness, and shortness of breath on slight exertion. About this time growths appeared on the temples and the sternocleido- mastoideus. Five months later the patient had three cramp-like attacks in one day, followed by great exhaustion. At this time the patient came to De Kenzi, who found small swellings over the entire body and great nervous de- pression. On the history given here he diagnosed cysticercus of the brain and skin. After a year's ambulant treatment with male fern the nervous trouble had disappeared, so had the swellings, with the exception of one over the left temple, and this was removed by operation and showed the presence of a cysticercus. The eosinophilia present at the beginning of the treatment had disappeared. 2. The second patient, a woman, had an infection with Twnia solium three years before. For two years previous to treatment she had suffered from dizziness, headache, weak memory, polyuria, and weakness. A swelling over the manubrium sterni showed all the claaracteristics of a cysticercus. After a week's treatment the nervous symptoms had disappeared and the swelling was reduced to one-third its original volume. A microscopic examination of the swelling was made, but De Renzi states that in this examination nothing of special importance was noted. 3. In De Renzi's third case there was a history of increasing pain in the hypochondrium for 20 months, no fever or emaciation, increase in the area of liver dullness, eosinophilia present, urine normal. The case was diagnosed as hepatic echinococcosis, and all symptoms disappeared under treatment in 20 days. * * * Fecal examination did not show parasites or their eggs. 4. The fourth patient was a woman who had suffered for a year with a pain in the thorax and often ctfughed blood. In the absence of tubercular symptoms and because the patient coughed up membranes, " Hiiutchen," a diagnosis of lung echinococcosis was made by De Renzi and confirmed by two associates. In the brief period of six days she was cured by male fern. Dianoux (1909) has recorded a case in which ocular and subcu- taneous cysticercosis, possibly complicated by cerebral cysticercosis, was apparently cured by the administration of male fern. Before coming under Dianoux's care, March 25, the patient had had several epileptiform attacks, and had nearly lost the sight of the left eye. Examination with the ophthf-lmoscope showed the presence of a cysticercus in the vitreous humor, and some weeks later a nodule, presumably a cysticercus, was found beneath the skin of the groin. After treatment with male fern and calomel the patient passed 4 or 5 meters of tapeworm {Twnia solium). May 2 the patient experienced an epileptifoim convulsion. From May 3 to 20 the patient took 2 grams of extract of male fern per day ; treatment was then suspended a few days. On May 24 a slight epileptiform attack occurred. Examination with the ophthalmoscope showed that the cysticercus had become shriveled and motion- less. From May 2.^5 to June 5, 2 grams of extract of male fern per day were administered. June 12 the cyst in the groin was discovered. June 16 an epi- leptiform attack occurred, lasting 15 minutes, and the following day there was another attack. Treatment was resumed. July 2 treatment was suspended; the cyst had disappeared from the gi-oin ; the general health of the patient was excellent. During 10 days in August the patient was given 1.5 grams of extract of male fern per day. November 15 the patient was discharged as cured. The cysticercus had entirely disappeared from the eye. The only evidence of its former presence was a cicatrice and vascular condition of the retina at the HISTORICAL REVIEW. 9 point where the parasite had been located. Altogether 102 grams of extract of male fern had been administered during a period of 71 days, ftnd presuniably as a result of this treatment a cysticercus of the eye, one under the skin of the groin, and probably others in the motor centers of the brain, had degenerated and become absorbed. Dianoux concludes that male fern destroys cysticerci because of some selective action against these parasites. Eailliet in his abstract of De Renzi's article (De Renzi, 1909) has noted that it would be interesting to test male fern on domestic animals suffering from parasitic diseases of the muscles and viscera, and mentions gid as one disease in which this treatment should be attempted. Such treatment was attempted and reported by Hall (1909). The treatment was tried on three sheep as follows: t^Kf'*^'^ 1. Sheep showed characteristic symptoms of gid. Fifty grams of male fern powder administered on two consecutive days. The third day the attendant accidentally got the dose in the windpipe of the sheep, killing the animal. Post-mortem examination showed a large living coenurus in the cerebrum. ( 2. Sheep showed pronounced symptoms of gid. Ethereal extract of male fern administered in 5 c. c. doses on 27 days between April 22 and May 30, a total of 135 c. c. Sheep found dead May 30, following a gradual increase in unfavorable symptoms from May 1. Post-mortem examination showed a large living coenurus in the cerebrum. 3. Sheep showed characteristic symptoms of gid. Ethereal extract of male fern administered in 5 c. c. doses on consecutive days, with the exception of one day, until a total of 45 c. c. had been given. No improvement in condi- tion. Treatment discontinued and sheep found dead four days later. Post- mortem examination of the brain showed a small live coenurus. Hall has given a critical review of De Renzi's cases, and concludes that they are open to suspicion of error as regards diagnosis and the connection between the disease, the treatment, and the cure. Moussu (1910) has also reported some tests of ethereal extract of male fern administered for two months to two giddy sheep. There was a marked amelioration of symptoms, but at the autopsy, two months after the cessation of treatment, there was a coenurus in the brain of each, very large in one case and the size of a hazelnut in the other, and neither of them degenerated. Moussu also tested the treatment on a pig affected with cysticercosis which had been devel- oping for more than six months, the vesicles being readily visible under the tongue and the conjunctiva. The condition of the pig, instead of improving, became worse from day to day. In view of the conflict between his results and those obtained by De Renzi and Dianoux, Moussu concludes that the influence of male fern only makes itself felt on young lesions and those in course of development, but that where the cysts are entirely developed the medicament remains entirely without effect. The most recent publislied work upon this subject available to us is that of Deve (1911). His test of the treatment was as follows: Three rabbits were given a snt)cutaneou8 injection of brood capsules from a iydatid cyst. No. 1 was kept as a check. No. 2 was given each day for 52 50CG4"'— Bull. 15.3—12 2 ^0 . ACTION OF ANTHELMINTICS ON PARASITES. days 4.5 c. c. of ethereal extract of male fern. The weight of the rabbit was 2.9 grams. No. 3, weighing the same, was given 9 c. c, or 6 c. c, per kilogram, or double the corresponding dose for man. The treatment was begun the same day as the injection. Five months later normal live echinococcus cysts were found in all three rabbits. From this Dev6 concludes that in doses as large or twice as large as De Reuzi used and extending over a much greater length of time the male fern was without effect on the hydatid cysts. The treatment showed itself incapable of affecting the delicate metamorphosis of the scolex, an initial phase in which it seems that the parasite would be especially vul- nerable. Male fern is therefore provisionally deemed inefficacious in hydatid disease. Deve notes that his results in echinococcosis are in agreement with the negative results obtained by Hall in cysticercosis (Deve's error; Hall's cases dealt with coenurus, as noted above), by Moussu in ccenurosis, and finally with the very unsatisfactory results ob- tained by Railliet, Moussu, and Henry in distomatiasis. A summary of the foregoing indicates that results following the administration of male fern were claimed as follows : Successful in 6 cases of human cysticercosis (4 subcutaneous, 1 subcutaneous and cerebral, and 1 subcutaneous and ocular), and in 2 cases of human echinococcosis (1 hepatic and 1 pulmonary) ; a total of 8 cases of Imman somatic tseniasis. Unsuccessful in 5 cases of ovine cerebral ccenurosis, 2 cases of leporine general echinococcosis, and 1 case of porcine muscular and ocular cysticercosis; a total of 8 cases of somatic tseniasis in lower animals. A critical discussion of these cases will be given later. It is suffi- cient at this point to note that all tests on man were claimed to be successful and that all experiments on animals were failures. The cases of Ziirn and Curtice are too indefinite to include in this sum- mary. DISTOMATIASIS. Under this heading hepatic distomatiasis will be considered first and venal distomatiasis second. Grassi and Calandruccio (1884 and 1885) record the first cases of which we are aware where extract of male fern was used in dis- tomatiasis. The article published in 1884 is not available to us, but from the article of the following year it appears that they ad- ministered to sheep a single dose consisting of 5 grams of ethereal extract of male fern in 50 grams of ethereal tincture of male fern. They .note that one might use injections of male fern, but that it would not always be necessary. The injection should consist of 1 gram of the ethereal extract mixed with 1 gram of the tincture and injected directly into the liver by means of a Pravaz syringe. A postscript, based on later work, adds that this injection should not HISTORICAL REVIEW. 11 be employed. They find that for 24 to 48 hours after the adminis- tration of tlie medicine the feces contained numerous flukes and nematodes. After three days the feces show no eggs, and on autopsy D£> flukes or strongyles are found. The number of tests and other details are not given. They point out that this remedy should be tried in human dis- tomatiasis, and that the injection method might be useful in echino- coccus infections. Perroncito (1886) has repeated the experiments of Grassi and Calandruccio and reports favorably on the use of male fern, noting, however, certain unfavorable results from its use. His experiments were as follows: 1. Sheep with symptoms of fluke disease. Fecal examination showed two to three eggs of Strongyliis, probably 8. cotitoi'tvs, and three to four fluke eggs per slide. Administered 10 grams of ethereal extract of male fern in 48 grams of ethereal tincture. The animal became tympanic, due to ethereal vapor de- veloped in the stomach, and in a few minutes fell as if struck dead. After 40 minutes rose. At the end of 2 hours it had recovered from the symptoms of anesthesia and other phenomena of etherization. Some weeks later the feces showed one strongyle egg and one distome egg per slide. 2. Administered to sheep affected with liver fluke, 5 grams of ethereal extract of male fern in 50 grams of the tincture. Usual symptoms of flatulence and defecation. After 30 to 40 minutes sheep gradually returned to normal condi- tion. Examination of the feces showed numerous distome and strongyle eggs. Two days later examination showed distome and hookworm eggs, but notice- ably diminished in number. There had been 10 to 12 eggs per slide ; there were now 4 to 5. Two days later repeated the dose, substituting 6 grams of ethereal extract in place of 5 grams. The animal showed more severe symptoms than on the first occasion. There was considerable flatulence at the end of an hour. Feces collected 40 hours after the second dose of male fern showed 2 to 3 distome eggs per slide. • Animal seemed much improved. Seven days after the second dose the feces showed only one distome egg to three or four slides. 3. A sheep infested with a large number of liver flukes, about 800, was given a large dose of male fern and died iu 10 or 12 hours. Autopsy showed that the flukes were apparently all dead. This last case had been previously published by Perroncito (1885). The next article dealing with experimental medication of hepatic distomatiasis which has come to our attention is that of Romagnoli (1903), who claims to have had good results in the treatment of dis- tomatiasis in sheep from the daily administration of 1 grain of salol in a teaspoonful of water for 8 days. According to that author the salol kills the cercarise which are still in the stomach and thus pre- vents further infestation, so that if the sheep are meanwhile given plenty of nourishment they rapidly recover their health and finally become completely cured. Romagnoli's treatment is of a different character from that reported by other writers in that it is supposed 12 ACTION OF ANTHELMINTICS ON PARASITES. to act upon jDarasites during their invasion of the body and not upon those which have alread}" become established in the host Floris (1907, 1908) has used carbon bisulphid, a remedy which Perroncito and Bosso (1894) and Wessel (1901) had reported as efficacious against Gastro'plnliis in the horse, and which Taar (1907) had found efficacious against Gastroph'dus and Ascaris in the horse. Floris administered the carbon bisulphid in gelatin capsules in doses of 10 to- 15 grams three or four times a week. The treatment was not attended by unpleasant effects and served to free the animals of flukes, the feces containing 5 to 10 flukes at a time. Floris notes that this is a very inexpensive remedy. Alessandrini (1908) reports that he had administered extract of male fern to two sheep which were severely infested. The sheep were dead two days later. Autopsy showed the flukes in the intestine and liver to be dead and degenerated. Pericaud (1908) has a rather glowing account of the virtues of a so-called " distomasine," said to consist of a glucosid and some plant essence. He gives no experimental tests, and the paper apparently adds nothing to our knowledge. Borini (1911) states that in 1905, at the suggestion of Perroncito, ethereal extract of male fern was used to arrest a plague of bovine distomatiasis occurring on the estate of a rich proprietor in Calabria. Experiments have been made on laboratory animals from that year on, the treatment being especially tested on sheep. These experi- ments, according to Borini, confirm tlie therapeutic value of male fern against distomatiasis. In light infections cures were always obtained; in the worst cases of advanced cachexia the treatment failed. The use of thymol in connection with male fern is recom- mended. Railliet, Moussu, and Henry (1911) have recently published a series of articles on the treatment of distomatiasis. In their first papers (1911a and 1911b) they note the desirability of some medica- tion in view of the considerable losses suffered in France in 1910. They first experimented with medicines^ Avhich maj?^ be eliminated by the liver, especially aloes, calomel, sodium salicylate, and "boldo." Most of the animals treated with these remedies improved, but there was no cure, no destruction of the parasites. Autopsy showed the flukes to be still alive after treatment extending over 15 days to three weeks. It seemed as though the treatment reduced the activity of the flukes without really having a specific effect on them. In another series of experiments they used compounds of phos- phorus, arsenic, and mercury-, as phosphorated oil, arsenic, atoxyl, arseno-benzol, trypanblau, benzoate of mercury, and fluid extract of broom. None of them gave certain, jjositive results. HISTORICAL REVIEW. 1^ In a third paper (Eailliet, Moussu, and Henry^ 1911c) these authors report better success in their tests of a new series of sub- stances, including particular^ tartar emetic, urotropin, atoxyl, and ethereal extract of male fern. The sheep used were for the most part heavily infested and had an intense cirrhosis and often a perforation of the capsule of Glis- son, the flukes which caused the perforation escaping to the peri- toneal cavity. Many had a mixed infection with Fasciola hepatica and Dicrocmlium lanceatum. Tartar emetic and urotropin proved to be inefficacious. Atoxyl produced the evacuation of degenerated F. hepatica in one heifer. The tests with ethereal extract of male fern were more satisfac- tory. An abstract is given of Alessandrini's (1908) experiments with male fern, noting this as the only work of the sort of which they were aware. Their own tests were made on five sheep, three others being kept as checks. The sheep were given 5 grams of ethereal extract in 25 c. c. of oil. Autopsy was made immediately after death. The experiments are as follows : 1. Sheep received one dose and died 12 hours later. Three hundred and ninety F. hepatica and numerous D. Janccatum in liver; all alive. 2. Sheep received two doses at 16-hour intervals and was killed in extremis 24 hours after the second dose. The biliary canals contained numerous live D. lanceatum and 55 F. hepatica, 4 of them being alive and the rest degen- erated and of a yellowish-green color. The gall bladder contained 142 F. hepatica, all dead, though only 3 showed the same alteration as the preceding. The small intestine contained 16 degenerated specimens and the large intestine contained 10 dead but not degenerated specimens. 3. Sheep received three doses at 16 and 24 hour intervals. Killed 3 days after the last dose. Biliary canals contained several live D. lanceatum. There were 9 F. hepatica, all degenerated, in the gall bladder, but none in the biliary canals or in the intestine. 4. Sheep received four doses at 24, 24, and 48 hour intervals. Died 7 hours after the last dose. Biliary canals contained numerous live D. lanceatum. There were 5 degenerated F. hepatica in the gall bladder, none in the biliary canal, 1 degenerated specimen in the large intestine, and about 50 live speci- mens in the peritoneal cavity, associated with peritonitis. 5. Sheep received four doses at 48, 24, and 24 hour intervals. Killed 3 days after the last dose. Biliary canals contained several D. lanceatum, all living. There were no /•'. hepatica in the bile ducts, the gall bladder, or the intestine. The three check sheep were killed at the end of the experiment and showed 178, 85, and 497 F. hepatica, respectively, and numerous D. lanceatum, all living. The degeneration undergone by F. hepatica under the influence of male fern, and to a lesser extent of atoxyl, begins at (he posterior extremity and progresses anteriorly, some Individuals being green and flabby at the posterior end, while the cephalic end was still capable of movement. They conclude that male fern is a satisfactory treatment for in- fection with F. hepatica if taken before there are irremediable lesions and at least four doses of 5 grams each administered. This 14 ACTION OF ANTHELMINTICS ON PARASITES. will kill F. hepatica in the liver, but not in the peritoneal cavity. At the same time it is effective against gastrointestinal strongylosis. It has no effect on D. larweatum. In a later paper (Railliet, Moussu, and Henry, 1911d) these authors recapitulate the foregoing, noting the earlier work of Grassi and Calandruccio and of Perroncito. They add the following experiments : Four sheep were given 5 grams of ethereal extract of male fern in 25 c. c. of oil for four successive days, a fifth animal being kept as check. All the sheep showed fluke eggs in the feces. Four days after the last dose they were all killed. Autopsy showed the following : 1. Liver contained 1 F. hepatica (presumably alive), but marked lesions of cirrhosis indicated the recent disappearance of other flukes. 2. Liver contained 2 live F. hepatica in the terminal ducts and 3 dead and degenerated forms. Hepatic lesions moderated. 3. Liver, abnormal in appearance, contained 26 live F. hepatica and 1 dead one. 4. Liver contained 1 live F. hepatica. 5. Check animal. Liver strongly cirrhotic, contained 296 live F. hepatica. All animals contained a number of live D. lanceatum. They conclude that male fern is effective, and, for sheep, suggest a dose of 1 gram of ethereal extract for every 5 kilos of live weight of animal ; for cattle, about 30 grams for 350 to 400 kilos of live weight. They note that male fern is comparatively cheap and suggest that it be administered by means of a rubber tube. The status of anthelmintics in bilharziasis is indicated in the following summary: Stiles (1904) says: "Favorable results are claimed from repeated doses of male fern ; some authors consider specific treatment futile." Sandwith (1909) writes of bilharziasis : No method has as yet been discovered of killing the worms in the human body; the ordinary vermifuges are useless. * * * The liquid extract of male fern, in doses of 15 minims 3 times a day, is the only drug of known value, for, though it does not expel the parasites [How could it?], it seems to weaken their power of doing harm ; it diminishes hematuria, allays vesical irritation, and reduces the number of eggs passed in the urine and feces. Joannides (1911) has tried salvarsan in bilharziasis in 8 cases, and reports that a single injection of 0.5 to 0.6 gram (?) of salvarsan in the case of adults and 0.25 gram (?) in the case of a 12-year-old child resulted in cessation or great diminution of hematuria, relief from vesical and urethral irritation, and a disappearance of the eggs from the urine. He concludes that salvarsan is destructive of Schis- tosomum hcematohium and its eggs and thus brings about a cure of the disease. Looss (1912) takes exception to the conclusions of Joannides and states that bilharziasis is a disease characterized by lesions due to the passage through or retention in the tissues of the bladder, rectum, HISTORICAL REVIEW. 15 etc., of vast numbers of eggs, the parasite itself in the blood vessels having practically no direct pathological significance. By the time the patient goes to a physician for treatment for hematuria the worms producing the eggs which cause this are probably dead or near their end, Looss looks upon the cures reported by Joannides as resulting from an artificial retention of the eggs in the tissues, causing thereby a suppression of the symptoms but not a cure of the disease. For the purpose of this article — a consideration of the question whether anthelmintics are effective against metazoan para- sites located elsewhere than in the digestive tract — the question as to whether the killing of the fluke is desirable or not, or as to whether the bilharzia disease is, strictly speaking, one due to flukes or to fluke eggs, is not material. Looss does not attack Joannides's con- clusions that the embryos in the fluke eggs and possibly the adult' flukes, also, are killed by salvarsan. Conor (1911) has tried salvarsan in one case of bilharziasis and found it inefficacious. Eggs containing living miracidia were found in the patient's urine every day but one for a month after the treatment. Fiillebom and Werner (1912) have also tried salvarsan in a case of bilharziasis, with the same negative result. Day and Richards (1912) have criticized Joannides's findings, and report three cases in which salvarsan was administered with no effect on the passage of living ova or, in two cases examined, on the eosinophilia. A summary of the foregoing papers on the treatment of hepatic distomatiasis shows the following: The administration of anthelmintics has been declared effective by Grassi and Calandruccio as a result of experiments (number not given) followed by fecal examination and post-mortem (male fern) ; by Perroncito after three experiment cases followed by fecal ex- amination and post-mortem (male fern) ; by Floris after experiments (number not given) in which no autopsies or subsequent fecal ex- aminations showing absence of eggs are noted (carbon bisulphid) ; by Alessandrini after two experiment cases followed by autopsy (male fern) ; by Borini after experiments (number not given) in which no autopsies or subsequent fecal examinations showing absence of eggs are noted (male fern and thymol) ; and by Railliet, Moussu, and Henry after nine experiment cases, four other animals being used as checks, followed by autopsy (male fern). This is a total of 14 detailed experiments and 2 other sets of tests with the number of animals not given. Romagnoli's treatment is omitted from con- sideration in this summary, as no claim has been made that it affects parasites already established in the host. In the case of venal distomatiasis, male fern has been commended as being* efficacious, with no data found by us on casual examination 16 ACTION OF ANTHELMINTICS ON PARASITES. of the literature as to autopsy showing that the death of the fluke actually follows the administration of the male fern. Salvarsan has been commended on the basis of a cessation of symptoms following its administration in eight cases of bilharziasis, and opposed on the find- ings in five cases in which its administration was without effect. Looss questions the benefits of salvarsan as regards the organic lesions due to tlie parasite, while the other writers take exception to Joannides's findings as not conforming to other knoAvn facts. AUTHORS' EXPERIMENTS. Being engaged in a study of tapewonn disease of sheep, due to Thysanosoma actinioides, the authors have taken advantage of oppor- tunities to test carbon bisulphid and male fern as remedies for this disease. At the request of a sheep owner we also tested a certain proprietar}'^ remedy. This remedy was found on analysis by the Biochemic Division of the Bureau of Animal Industry to consist of 49 per cent ferrous sulphate, 13 per cent arsenious oxid, 8 per cent oxids of calcium, potassiimi, silicon, and magnesium^ and 29 per cent organic matter, nature not determined but not containing santonin, or any other vegetable alkaloid. Tests of carbon bisulphid were made by the junior author at the ranch of Mr. Wells, near Resolis^ Colo., in July, 1911. A Mexican, herder at one camp was instructed to cut out six sheep, picking the poorest and weakest as being the most likely victims of thysano- somiasis. Events proved that some of the sheep selected were really" too sick for the experiment. The experiments were as follows: 1. Administered 6 grams of carbou bisulphid in capsule to a sheep. Sheep crushed one or two capsules and seemed greatly distressed thereby. Repeated- dose after 30-hour interval. Sheep found dead the next day. Post-mortem showed appearances similar to those of hemorrhagic septicemia. A number of live bile-staiuetl Thysanosom