Pathology in hamsters administered Solanum plant species that contain steroidal alkaloids.

Syrian hamsters were orally administered ground plant material from either Solanum sarrachoides, S. melongena, S. eleagnifolium, or S. dulcamara. Six of eight hamsters administered S. eleagnifolium and eight of 10 hamsters administered S. dulcamara died following administration of plant material and had gastric glandular mucosal necrosis and small intestinal mucosal necrosis with little inflammation. Hamsters administered S. sarrachoides or S. melongena did not die and had only lesions compatible with gastric distension. Both S. eleagnifolium and S. dulcamara contained solasodine glycoalkaloids(s), and S. dulcamara also contained an equal amount of other glycoalkaloids which were probably derived from soladulcidine (dihydrosolasodine). The lesions produced by these two plants were similar to those reported earlier to be caused by sprout material of S. tuberosum (in which solanidane alkaloids predominate) and by an alkaloid extract of S. tuberosum sprouts. Because of similarities in saponin-like activity and structure of solasodine glycoalkaloids to the solanidine glycoalkaloids of potato sprouts, the glycoalkaloids of S. dulcamara and S. eleagnifolium were probably the agents responsible for the lesions observed.

Spirosolane-containing Solanum species and induction of congenital craniofacial malformations.

Comparison by GC analysis of purified alkaloid extracts of Solanum species revealed no measurable free solasodine, other spirosolanes, or any non-spirosolane steroidal alkaloid aglycones in unhydrolyzed total alkaloid fractions of fruit of Solanum elaeagnifolium Cav. (silverleaf nightshade), Solanum sarrachoides (S. villosum Lam.--hairy nightshade), Solanum dulcamara L. (European bittersweet nightshade) or Solanum melongena L. (eggplant). All alkaloidal material was apparently present as glycoside. Conversely, sprouts of Solanum tuberosum L. (potato) contained 67% of its alkaloids as glycosides, which was freed only upon hydrolysis with the remaining 33% present as free solanidine. GC/MS analysis of hydrolysates of purified extracts of the test Solanum species revealed that solasodine was a principal or sole aglycone of the alkaloid glycosides in each of the test species except Solanum tuberosum. In the latter, solanidine was the sole aglycone. Among the test species, exclusive of S. tuberosum, only S. dulcamara contained aglycones other than solasodine. In addition to solasodine, S. dulcamara contained appreciable amounts of an unknown spirosolane, an aglycone provisionally identified as soladulcidine. The induction of congenital craniofacial malformations in hamsters by high oral doses of the four Solanum species that contained mainly solasodine glycosides--S. elaeagnifolium, S. dulcamara, S. sarrachoides and S. melongena was compared to inductions of malformations by Solanum tuberosum, that contained mainly solanidane glycosides. Compared to controls, Solanum elaeagnifolium and Solanum dulcamara fruit both induced a high percentage incidence of deformed litters (20.4 and 16.3, respectively) that was statistically significant (P less than 0.001 level) while percentage incidence of deformed litters induced by Solanum sarrachoides and Solanum melongena fruit (9.5 and 7.6 respectively) were both higher than controls (3.4%), in neither case was the incidence statistically significant (P less than .05). Deformed litter incidence induced by sprouts of Solanum tuberosum was 24.0%, (P less than 0.001).

Congenital skeletal malformations and cleft palate induced in goats by ingestion of Lupinus, Conium and Nicotiana species.

Three piperidine alkaloid containing plants, Conium maculatum (poison-hemlock), Nicotiana glauca (tree tobacco) and Lupinus formosus (lunara lupine), induced multiple congenital contractures (MCC) and palatoschisis in goat kids when their dams were gavaged with the plant during gestation days 30-60. The skeletal abnormalities included fixed extension or flexure of the carpal, tarsal, and fetlock joints, scoliosis, lordosis, torticollis and rib cage abnormalities. Clinical signs of toxicity included those reported in sheep, cattle and pigs--ataxia, incoordination, muscular weakness, prostration and death. One quinolizidine alkaloid containing plant, Lupinus caudatus (tailcup lupine), on the other hand, which is also known to cause MCC in cows, caused only slight signs of toxicity in pregnant goats and no teratogenic effects in their offspring.

Concentration of galegine in Verbesina encelioides and Galega oficinalis and the toxic and pathologic effects induced by the plants.

Verbesina encelioides administered to sheep by gavage induced clinical signs of toxicity and pathologic lesions identical to those induced by Galega officinalis. Sheep had compromised respiratory function with shallow, rapid respiration and frothy exudate from the nares. Affected animals necropsied at time of death presented with hydrothorax with as much as 2 to 3 L of straw-colored thoracic fluid with fibrin tags and congestion and edema of the lungs. The trachea and lung airways contained frothy material with fibrin strands. In some cases, subendocardial hemorrhage of the left ventricle was present. Galegine, a guanidine compound believed to be responsible for these effects, was found at an average concentration of about 0.46% in Galega and at 0.08% in the Verbesina collection that induced toxicosis. While G. officinalis is a known poisonous plant, its very limited distribution in the U.S. causes it to be of minor importance. V. encelioides, however, is widely distributed in the U.S. and presents a potential hazard for grazing livestock. Verbesina may have been responsible for past livestock deaths in the U.S., and thus should be classified as a poisonous plant.

Cyclopamine and related steroidal alkaloid teratogens: their occurrence, structural relationship, and biologic effects.

A spontaneous congenital deformity is produced in lambs whose dams consume Veratrum californicum on the 14th day of gestation. The deformity is generally expressed as cyclopia, cebocephaly, anophthalmia, or microphthalmia. This teratogenic effect is produced by certain steroidal alkaloid teratogens from the plant - most notably the compound cyclopamine. Cyclopamine is a C-nor-D-homo steroid with fused furanopiperidine rings E and F at right angles to the plane of the steroid because of spiro attachment at C-17 of the steroid. Among veratrum alkaloids, only those with an intact furan ring E were teratogenic in sheep, whereas those in which the peperidine ring is not rigidly positioned at right angles to the steroid were not. Many ruminants and laboratory animals are susceptible to the teratogen. It has wide species and tissue specificity and appears to have a direct effect on the embryo, not as a consequence of metabolic alteration of its structure nor as an indirect effect through a maternal influence. Other plant sources, notably potatoes, tomatoes, and eggplant contain related spirosolane steroidal alkaloids. Among naturally occurring spirosolanes, solasodine is teratogenic in hamsters, but neither tomatidine not diosgenin, the non-nitrogen containing analog of solasodine, is teratogenic. Results of these and other studies suggest that a basic nitrogen positioned alpha with respect to the steroidal plane and at appropriate distance beyond the D ring confers the teratogenicity on the molecule. Potato sprouts with high alkaloid content are teratogenic in hamsters, but tubers and peels are not.

Structure and stereochemistry of steroidal amine teratogens.

Studies of teratogenic steroidal alkaloids from Veratrum and Solanum have shown that those bearing a basic nitrogen atom in ring F, shared or unshared with ring E, with bonding capabilities alpha to the steroid plane may be suspect as teratogens. Examples of steroidal alkaloids which produce terata but, until recently, have been of uncertain structure include muldamine and the isomeric 3, N-diformylsolasodines. The correlation of their structures with the structure-terata relationship developed by Keeler and Brown is discussed. A brief introduction to teratogenicity is presented.

Myopathy in cattle induced by alkaloid extracts from Thermopsis montanta, Laburnum anagyroides and a Lupinus sp.

A purified alkaloid preparation containing N-methylcytisine, cytisine, 5,6-dehydrolupanine, thermopsine and anagyrine from Thermopsis montana induced prolonged recumbency and microscopic acute hyaline skeletal myodegeneration with myofibre regeneration in cattle similar in type and severity to that induced by Thermopsis montanta plant material. This indicates that the alkaloid(s) of Thermopsis montana are responsible for the myopathy caused by the plant. An alkaloid preparation containing mostly anagyrine from a Lupinus sp. and an alkaloid preparation containing only cytisine from Laburnum anagyroides each caused microscopic skeletal muscle degeneration and necrosis similar to, but less severe than, the alkaloid extract from T. montana, but without clinical recumbency. Dosage and severity of response suggest that neither of those two alkaloids alone can account for the effects induced by Thermopsis. The data suggest that quinolizidine alkaloids with a alpha-pyridone A-ring may be responsible for the lesions and that individual alpha-pyridones may have additive effects.

Livestock models of human birth defects, reviewed in relation to poisonous plants.

Certain birth defects in livestock induced by poisonous plants possess clinical similarities to congenital deformities in humans. They meet some criteria as models for those corresponding human conditions. They are induced by several plants, including members of the Astragalus, Lupinus, Conium, Nicotiana and Veratrum genera. The terata expressions include effects in bone and soft tissue, particularly in the limbs, spinal cord and cephalic regions. Whether these livestock conditions become extensively used as models of human terata will be determined by how well they meet the criteria sought in good models. These prospective models have many favorable characteristics; however, being large domestic livestock, they have certain logistic disadvantages compared with small rodents as models.

Teratogens in plants.

Many compounds synthesized by plants are known to be teratogenic in laboratory animals, but only a few have been shown by feeding trials to produce terata in livestock. Studies of plant teratogens affecting livestock have not moved forward in a systematic nor rapid way because of the logistical problems connected with such experiments in large animals. Information that has accumulated can be conveniently separated into three categories: (1) known teratogens in known teratogenic plants, (2) known teratogenic plants with unidentified teratogens, and (3) suspected teratogenic plants. Included in the first group are the teratogens from Lupinus, Veratrum, Conium and Leucaena genera; in the second group are included the Astragalus , Nicotiana and Trachymene genera; and in the third group are included Datura, Prunus , Sorghum and Senecio genera. Total available information in each case varies, but in a few instances considerable fundamental as well as practical information is now known. Research has provided enough information in a few instances to enable elimination of the practical problem or significant reduction in incidence.

Toxicoses in livestock from the hemlocks (Conium and Cicuta spp.).

The hemlocks, Conium maculatum (poison-hemlock) and Cicuta spp. (waterhemlock), are poisonous plants that cause sizeable losss to the livestock industry. Clinical signs of poisonhemlock toxicosis are similar in all species of livestock and include muscular weakness, incordination, trembling, initial central nervous system stimulation, depression and death from respiratory paralysis. Poison-hemlock also causes skeletal defects in the offspring of cattle, pigs and sheep and cleft palate in pigs when ingested during specific periods of gestation. The primary toxicants in poison-hemlock are coniine and gamma-coniceine. Coniine predominates in mature plants and seed, whereas gamma-coniceine predominates in early growth of the plant. Waterhemlock is the most violently toxic poisonous plant known. The toxicant is cicutoxin, which acts on the central nervous system, causing violent convulsions and death. Clinical signs of poisoning appear within 15 min after ingestion of a lethal dose and include excessive salivation, nervousness, tremors, muscular weakness and convulsive seizures interspersed by intermittent periods of relaxation and a final paralytic seizure resulting in anoxia and death. Elevated activities of lactic dehydrogenase, aspartate aminotransferase and creatine kinase in blood are observed, indicative of muscular damage. Toxicoses from poisonhemlock and waterhemlock generally occur in early spring when both plants emerge before other, more palatable plants begin to grow. All parts of the poison-hemlock plant are toxic. The root or tubers of waterhemlock are toxic; however, experimental evidence concerning the toxicity of other plant parts is inconclusive.

Maternal and fetal toxicity of poison hemlock (Conium maculatum) in sheep.

Poison hemlock (Conium maculatum) was toxic to pregnant ewes and their fetuses when fed during gestation days 30 through 60. Maternal effects included trembling, muscular weakness in the neck initially, then progressing to the limbs, ataxia, frequent urination and defecation, and death. Convulsive seizures were not observed. Fetotoxic effects included excessive flexure of the carpal joints with lateral deviation in the front limbs and kinked tails. At term, 7 of 11 lambs had varying degrees of the limb abnormalities, but all lambs appeared clinically normal at 8 weeks after parturition.

Induction of cleft palate in newborn pigs by maternal ingestion of poison hemlock (Conium maculatum).

Cleft palates were induced in newborn pigs of gilts fed Conium maculatum seed or plant during gestation days 30 through 45. Twelve of 23 newborn pigs born to 3 gilts given Utah-grown C maculatum seed and 9 of 12 newborn pigs born to a single gilt given the fresh Utah spring-growth C maculatum plant had cleft palates. The cleft palates ranged from a unilateral cleft, involving only 1 side of the palate, to a full bilateral cleft. Brachygnathia was also observed in some of these newborn pigs with cleft palate. Other malformations were not observed. Chemical analysis of seed and plant samples indicated that gamma-coniceine was the responsible teratogenic alkaloid. A daily dose of plant or seed that provided greater than or equal to 1.07 mg of gamma-coniceine/kg of body weight fed to gilts during the 30th through the 45th day of pregnancy resulted in teratogenic effects.

Congenital skeletal malformations induced by maternal ingestion of Conium maculatum (poison hemlock) in newborn pigs.

Skeletal malformations were induced in newborn pigs from gilts fed Conium maculatum seed or plant during gestation days 43 through 53 and 51 through 61. The teratogenic effects in groups dosed during gestation days 43 through 53 were more severe than those in groups dosed during the later period, with many newborn pigs showing arthrogryposis and twisted and malaligned bones in the limbs and with 1 pig showing scoliosis and deformity of the thoracic cage. The pigs born to gilts given C maculatum during gestation days 51 through 61 had excessive flexure primarily in the carpal joints, without scoliosis or bone malalignment in the limbs. The teratogenicity of poison hemlock depends on the alkaloid concentration and content. Based on the data presented, we speculate that gamma-coniceine is the teratogenic alkaloid in the poison hemlock fed to the gilts.

Nicotiana glauca-induced congenital deformities in calves: clinical and pathologic aspects.

Seven calves born to 7 cows fed Nicotiana glauca during portions of the 1st trimester of gestation were deformed at birth. Deformities increased in severity as the calves aged. At birth, calves typically had arthrogryposis of the forelimbs or curvature of the spine; severity varied among calves. In 4 calves necropsied at about 15 months of age, there was general malpositioning and misalignment of the distal ends of the radius and ulna and the proximal ends of the metacarpal bones. Carpal joints were severely affected, fetlock joints were moderately affected, and pastern joints were slightly affected, with lateral rotation of forelimbs common; severity varied among calves. In 1 of the 4 calves, there was moderate torticollis and scoliosis resulting from wedging of some of the cervical and thoracic vertebrae and abnormal cranial curvature of the left thoracic ribs. Histologic changes were not noticed in muscle, brain, spinal cord, or endocrine organs of the 4 calves.

Thermopsis montana-induced myopathy in calves.

Six heifer calves were administered the dried and ground whole plant of Thermopsis montana by gavage once daily for 2 to 4 days at an initial dosage rate of 1 g/kg of body weight. Two of the 6 heifers died after the second dose, and the remaining 4 calves were humanely killed and their tissues were examined. Serum creatine kinase and aspartate transaminase activities were significantly (P less than 0.05) increased after initiation of plant administration. All calves had skeletal muscular degeneration or evidence of regeneration and repair of damaged myofibers in all skeletal muscle groups examined. Myoglobinuria and cardiac muscle degeneration were not detected in any of the calves.

Quinolizidine and piperidine alkaloid teratogens from poisonous plants and their mechanism of action in animals.

Quinolizidine and piperidine alkaloid teratogens from Lupinus, Conium, and Nicotiana genera have been identified as causes of birth defects in livestock induced by poisonous plants. Many defects now known to be related to poisonous plant ingestion were once thought to have a genetic origin. This supposition delayed diagnosis, reporting, and understanding of such birth defects, because breeders and producers feared the news would make it difficult to sell breeding stock. Defects caused by quinolizidine and piperidine teratogens include cleft palate and contracture-type skeletal defects such as arthrogryposis, scoliosis, torticollis, and kyphosis. Teratogens have been identified, differences in susceptibility to teratogenic compounds among livestock species have been elucidated, periods of gestation when specific types of birth defects occur have been determined, and information about mechanism of action has been developed.