[CANCER RESEARCH 28, 2323-2326, November 1968] Effects of Teratogenic Agents in Range Plants Wayne Binns, Lynn F. James, Richard F. Keeler, and Lew Dell Balls Agricultural Research Service, USDA, Animal Disease and Parasite Research Division, Poisonous Plant Research Laboratory, Logan, Utah 84321 SUMMARY commonly occurred was used for bioassays of the suspected plants. Each plant was mixed with water and fed daily to the A cyclopian-type malformation in lambs has been produced animals at a subclinical toxic level via stomach tube. This by maternal ingestion of Veratrum califomicum. Arthrogrypo- feeding was continued for a specific period during gestation. In sis, torticollis, scoliosis, and cleft palate in calves resulted from addition, experimental animals are fed alfalfa hay and mineral the maternal ingestion of two toxic species of lupine. Con supplements free choice. tracted tendons of the forelimbs and hypermobility of the hock and stifle joints were noted in lambs from ewes that had eaten locoweeds. RESULTS Two teratogenic steroidal alkaloids have been specifically A congenital cyclopian-type malformation in lambs was identified in V. califomicum An apparent relationship be found to be caused by maternal ingestion of V. califomicum tween locoism and lathyrism has been hypothesized on the (Fig. 1) with the time of insult being on the 14th day of basis of the botanical and chemical relationship of plants and gestation (1—3). The deformities ranged from anophthalmia, the similarity of congenital deformities. cyclopia, and/or cebocephalus, to a slight shortening of the nasal bones (Fig. 2). Prolonged gestation is commonly associ INTRODUCTION ated with the more severe deformities. The gestation period has been prolonged to as much as 87 days beyond the normal Numerous poisonous plants are present on the western gestation time of 147 to 152 days. If these prolonged gesta livestock range areas of the United States. They were not tions are interrupted by caesarian section or by a sacrificing of considered to be possible causative agents for congenital the ewe, the lambs are obtained alive, but die immediately anomalies in calves and lambs until 1963. At that time, it was from suffocation due to a bony partition in the nasophar- reported (1-3) that the maternal ingestion of Veratrum yngeal region obstructing the passage of air. califomicum by pregnant ewes on a specific day of gestation Maternal feeding of Lupinus sericeus or L. caudatus from the induced a cyclopian-type malformation in lambs. The terato 40th to the 70th day of gestation produced "crooked calf genic compound in V. califomicum that induces the cyclopi disease" which is common in all western states and Alaska (13, an-type deformity in lambs was subsequently isolated (6). 14). This period of insult caused a marked arthrogryposis of Other poisonous range plants also have been extensively the front legs (Figs. 3, 4). The deformities of torticollis, studied by research scientists of the Agricultural Research scoliosis, and cleft palate are apparently associated with other Service, USDA, Animal Disease and Parasite Research Division, insult periods. Poisonous Plant Research Laboratory, Logan, Utah. A com Locoweeds have been studied for many years by numerous mon congenital deformity, identified as "crooked calf disease" research scientists, but the USDA workers were the first to characterized by arthrogryposis and/or torticollis, scoliosis, report that maternal ingestion of locoweeds produced congeni and occasionally cleft palate (13, 14) has been known to be tal deformities in lambs. The deformities have ranged from a caused by maternal ingestion of Lupinus sericeus and L. slight and temporary to a marked and permanent flexing of caudatus. Locoweeds, Astragalus lentiginosus, A. pubentissi- the carpal or pastern joints and/or anterior flexure with mus, and Oxytropis sericeus have been shown to contain hypermobility of the hock joint (5). teratogenic agents that produce congenital anomalies in calves The teratogenic agents in V. califomicum have been identi and lambs by maternal ingestion (4, 5). fied (4, 6, 11, 12) as steroidal alkaloids. One, cyclopamine (8, 10), was found to be responsible for cyclopian-type deformi MATERIALS AND METHODS ties. Another alkaloid of similar structure (10), veratramine, induced leg deformities ranging from a slight lateral or medial In the V. califomicum work, from i to 1 ton of individual bowing of the front legs to marked flexure of the carpal joint species of possibly causative poisonous plants were collected and/or hypermobility of the hock and stifle joints. In some from range areas where specific types of congenital deformities cases, complete lack of muscular control was produced (7, 9). had been endemic. The plants were taken to the Poisonous A possible relationship between locoism and lathyrism (4, Plant Research Laboratory, Logan, Utah, air dried, ground, 11, 12) was hypothesized from the botanical relationship of and stored in plastic bags at 5°C. As is done in all our the plants, congenital deformities induced, and the chemical investigations, the species of animals in which the deformity characteristics. Preliminary assay of locoweed extracts suggests NOVEMBER 1968 2323 Downloaded from cancerres.aacrjournals.org on September 28, 2021. © 1968 American Association for Cancer Research. Wayne Binns, Lynn F. James, Richard F. Keeler, and Lew Dell Balls the presence of lathyrogens, aminoacetonitrile, and a, (Durand). IV. First Isolation of Veratramine and Alkaloid Q and a •y-diaminobutyricacid in locoweeds. Reliable Method for Isolation of Cyclopamine. Phytochemistry, 7: 303-306,1968. DISCUSSION 7. Keeler, R. F., Binns, W. Chemical Compounds of Veratrum californicum Related to Congenital Ovine Cyclopian Malforma The experimental feeding of poisonous plants and purified tions: Extraction of Active Material (29178). Proc. Soc. Exptl. teratogenic agents induced deformities identical to the respec Biol. Med., 116: 123-127,1964. tive congenital anomalies observed under range conditions. 8. Keeler, R. F., and Binns, W. Teratogenic Compounds of Veratrum Many poisonous and nonpoisonous plants may contain terato- californicum (Durand). II. Production of Ovine Fetal Cyclopia by Fractions and Alkaloid Preparations. Can. J. Biochem., Physical, gens only at specific stages of growth or when exposed to 44: 829-838, 1966. adverse environmental conditions. 9. Keeler, R. F., and Binns, W. Possible Teratogenic Effects of Veratramine (31639). Proc. Soc. Exptl. Biol. Med., 123: 921-923, REFERENCES 1966. 1. Binns, W., Thacker, E. J., James, L. F., and Huffman, W. T. A. 10. Keeler, R. F., and Binns, W. Teratogenic Compounds of Veratrum Congenital Cyclopian-Type Malformation in Lambs. J. Am. Vet. californicum (Durand). I. Preparation and Characterization of Med. Assoc., 46: 180-183, 1959. Fractions and Alkaloids for Biologic Testing. Can. J. Biochem., 2. Binns, W., James, L. F., Shupe, J. L., and Everett, George. A Physiol., 44: 819-828, 1966. Congenital Cyclopian-Type Malformation in Lambs Induced by 11. Keeler, R. F., and Binns, W. Teratogenic Compounds of Veratrum Maternal Ingestion of a Range Plant, Veratrum californicum. Am. californicum (Durand). III. Malformations of the Veratramine- J. Vet. Res., 24: 1164-1175, 1963. Induced Type from Ingestion of Plant or Roots (32474). Proc. 3. Binns, W., Shupe, J. L., Keeler, R. F., and James, L. F. Chronologic Soc. Exptl. Biol. Med., 726: 452-454, 1967. Evaluation of Teratogenicity in Sheep Fed Veratrum californicum. 12. Keeler, R. F., James, L. F., Binns, W., and Shupe, J. L. An Am. Vet. Med. Assoc., 147: 839-842,1965. Apparent Relationship Between Locoism and Lathyrism. Can. J. 4. James, L. F., and Binns, W. Blood Changes Associated with Comp. Med. Vet. Sci., 31: 334-341, 1967. Locoweed Poisoning. Am. J. Vet. Res., 28: 1107-1110,1967. 13. Shupe, J. L., James, L. F., and Binns, W. Observations on Crooked 5. James, L. F., Shupe, J. L., Binns, W., and Keeler, R. F. Abortive Calf Disease. Am. Vet. Med. Assoc., 151: 191-197,1967. and Teratogenic Effects of Locoweed on Sheep and Cattle. Am. J. 14. Shupe, J. L., James, L. F., Binns, W., and Keeler, R. F. Lupine, a Vet. Res., 28: 1379-1388,1967. Cause of Crooked Calf Disease. Am. Vet. Med. Assoc., 757: 6. Keeler, R. F. Teratogenic Compounds of Veratrum californicum 198-203, 1967. 2324 CANCER RESEARCH VOL. 28 Downloaded from cancerres.aacrjournals.org on September 28, 2021. © 1968 American Association for Cancer Research. Teratogenic Agents in Range Plants A Fig. 1. Veratrum californicum in flowering stage. Fig. 2. Cyclopian malformed lamb induced by maternal ingestion of Veratrum californicum. NOVEMBER 1968 2325 Downloaded from cancerres.aacrjournals.org on September 28, 2021. © 1968 American Association for Cancer Research. Wayne Binns, Lynn F. James, Richard F. Keeler, and Lew Dell Balls Fig. 3. Lupinas sericeus. Fig. 4. Crooked calf disease induced by maternal ingestion of Lupinus sericeus. 2326 CANCER RESEARCH VOL. 28 Downloaded from cancerres.aacrjournals.org on September 28, 2021. © 1968 American Association for Cancer Research. Effects of Teratogenic Agents in Range Plants Wayne Binns, Lynn F. James, Richard F. Keeler, et al. Cancer Res 1968;28:2323-2326. Updated version Access the most recent version of this article at: http://cancerres.aacrjournals.org/content/28/11/2323 E-mail alerts Sign up to receive free email-alerts related to this article or journal. Reprints and To order reprints of this article or to subscribe to the journal, contact the AACR Publications Subscriptions Department at [email protected]. Permissions To request permission to re-use all or part of this article, use this link http://cancerres.aacrjournals.org/content/28/11/2323. Click on "Request Permissions" which will take you to the Copyright Clearance Center's (CCC) Rightslink site. Downloaded from cancerres.aacrjournals.org on September 28, 2021. © 1968 American Association for Cancer Research. .