Pyridoxine (a vitamin B6) and its derivative pyridoxal as treatment for Albizia versicolor poisoning in guinea-pigs.

In the course of three experiments it was established that all the toxic effects of a lethal dose of Albizia versicolor pods (greater than 4.5 g/kg) in guinea-pigs could be countered by concurrent subcutaneous injection of pyridoxine (10 mg/kg). This treatment was also successful once severe symptoms had set in. Pyridoxal, on the other hand, was found to be ineffective as a therapeutic agent. The fact that pyridoxal does not counter the action of the toxin indicates an atypical site of action by the toxin as regards the normal pathways which require vitamin B6 as a co-factor.

Isolation of the toxin responsible for gousiekte, a plant-induced cardiomyopathy of ruminants in southern Africa.

Gousiekte was induced in sheep and goats by the oral and intravenous (i/v) administration of methanolic and ethanolic precipitates of aqueous extracts of Pavetta harborii. Further purification of the toxic principle was carried out by dialysis and ion-exchange chromatography. The toxin was eluted from the cation-exchange resin with 1 mol/l of ammonium acetate and finally purified on Sephadex G-10 and silica gel. Spots on thin-layer chromatographic plates (TLC) were visualized with ninhydrin. Gousiekte was induced with the pure toxin after i/v administration in goats. The yield of pure toxin was about 10 mg/kg of dried leaves.

Experimental Albizia versicolor poisoning in sheep and its successful treatment with pyridoxine hydrochloride.

Five sheep developed severe nervous signs after being drenched with Albizia versicolor pod-material. Four of these sheep were treated with pyridoxine hydrochloride (a vitamin B6) when the symptoms of toxicity became life-threatening. All the treated sheep recovered dramatically and completely after treatment while the untreated one died 2 h after receiving pod-material. A therapeutic dose of 20-25 mg pyridoxine hydrochloride/kg body mass given twice with an 8 h interval is the recommended treatment regimen. The route of administration will depend on the severity of symptoms. Chemical pathology and post-mortem findings are discussed.

Photosensitivity in South Africa. VIII. Ovine metabolism of Tribulus terrestris saponins during experimentally induced geeldikkop.

Geeldikkop was induced in a sheep by dosing it orally with a crude extract of the steroidal saponins from Tribulus terrestris. GC-MS analysis of the sheep's ruminal contents, bile, faeces and urine for free and conjugated sapogenins, revealed the general features of the metabolic pathway by which diosgenin and yamogenin glycosides were converted into the glucuronides of epismilagenin and episarsasapogenin, the major constituents of the biliary crystals that usually form during geeldikkop. Other steroidal saponins in the T. terrestris extract, including those derived from tigogenin, neotigogenin, gitogenin and neogitogenin appear to be non-lithogenic. The implications of these findings are discussed.

Photosensitivity in South Africa. VII. Chemical composition of biliary crystals from a sheep with experimentally induced geeldikkop.

Geeldikkop was induced in a sheep by oral administration of crude saponins from Tribulus terrestris. Centrifugation of the bile from this sheep gave a pale green sediment of crystalloid material which was insoluble in common organic solvents, but soluble in acetic acid. Analysis of the crystalloid material by 1H and 13C NMR, EDXA, TLC, LSIMS, and by acidic hydrolysis followed by TLC and GC-MS, revealed it to be composed principally of a 6:1 mixture of the calcium salts of the beta-D-glucuronides of the steroidal sapogenins epismilagenin and episarsasapogenin. The administered saponin was found to contain glycosides of the steroidal sapogenins diosgenin, yamogenin, epismilagenin, tigogenin, neotigogenin, gitogenin and neogitogenin in the ratio 10:7:1:11:7:35:25. A metabolic pathway for the conversion of diosgenin and yamogenin saponins to the biliary glucuronides is proposed.

Photosensitivity in South Africa. VI. The experimental induction of geeldikkop in sheep with crude steroidal saponins from Tribulus terrestris.

Geeldikkop was induced in sheep by the oral administration of crude steroidal saponins from Tribulus terrestris. Two of the sheep developed typical lesions of geeldikkop, including birefringent crystalloid material in bile ducts and concentric periductal lamellar fibrosis. The clinical pathological changes in these sheep were also consistent with those of geeldikkop: aspartate transaminase and gamma-glutamyl transferase activities in the sera of both were elevated, and one had bilirubinaemia. A third sheep became photosensitive without typical lesions of geeldikkop in the liver or changes in the activities of liver enzymes before euthanasia. The findings of these trials are consistent with reports from abroad that ovine hepatogenous photosensitization, caused by Agave lechuguilla and Narthecium ossifragum, can be induced with crude saponins from the respective plants.

Krimpsiekte in a sheep following a single dose of Tylecodon ventricosus (Burm. f.) Toelken and the isolation of tyledoside D from this plant species.

Tylecodon ventricosus induced severe respiratory distress in two penned sheep without any electrocardiographic abnormalities being recorded. Based on the results it appears as if T. ventricosus predominantly induces the neuromuscular syndrome referred to as krimpsiekte. A single, relatively large intraruminal dose of 10.0 g/kg induced krimpsiekte in one sheep. Treatment with 5.0 g/kg activated charcoal on two consecutive days did not prevent the development of krimpsiekte. A bufadienolide, tyledoside D, was isolated from semi-dried plant material.

Krimpsiekte, associated with thalamic lesions, induced by the neurotoxic cardiac glycoside, cotyledoside, isolated from Tylecodon wallichii (Harv.) Toelken subsp. wallichii.

The specific neurotoxic principle of Tylecodon wallichii (Harv.) Toelken subsp. wallichii, the cause of krimpsiekte in small stock, was isolated and identified as the previously described cumulative bufadienolide, cotyledoside. Krimpsiekte was experimentally induced in two sheep by the repeated intravenous administration of cotyledoside at the rate of 0.01-0.015 mg/kg body mass. On day 9, both animals developed clinical signs typical of krimpsiekte, which is characterized by tremors, paresis and recumbency. Both sheep had difficulty in controlling their hindquarters when attempting to lie down. No significant electrocardiograph abnormalities were detected during the experiment which confirms that cotyledoside at low doses does not overtly affect the electrical activity of the heart. No gross lesions were observed in the sheep. The most significant microscopic lesions comprised mild brain oedema and pronounced vacuolation of the white matter of thalamic nuclei. These lesions might explain some of the motor function deficiencies clinically observed in this syndrome. The previously held contention that these neurotoxic cardiac glycosides are indeed the cause of krimpsiekte is, therefore, confirmed.

Photosensitivity in South Africa. IX. Structure elucidation of a beta-glucosidase-treated saponin from Tribulus terrestris, and the identification of saponin chemotypes of South African T. terrestris.

Saponin C, a beta-glucosidase-treated saponin isolated from ethanol-water extracts of a South African collection of Tribulus terrestris, was shown by one- and two-dimensional NMR spectroscopy to be ruscogenin 1-O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-6)-acetylglucopyranoside++ +. GC-MS analysis of the hydrolysed ethanol-water (4:1) extracts of T.terrestris specimens from two of four sites, revealed high levels of ruscogenin and potentially lithogenic diosgenin saponins. Specimens from two other sites contained non-lithogenic saponins derived predominantly from tigogenin, neotigogenin, gitogenin and neo-gitogenin.

Krimpsiekte and acute cardiac glycoside poisoning in sheep caused by bufadienolides from the plant Kalanchoe lanceolata Forsk.

Three toxic bufadienolides , one characterized as hellibrigenin 3-acetate, have been isolated from Kalanchoe lanceolata Forsk. Typical signs of cardiac glycoside poisoning, involving the gastro-intestinal, neuromuscular and cardiovascular systems, could be induced by drenching the milled plant to sheep. Such signs could also be induced by dosing the bufadienolides to sheep or by injecting them into both guinea-pigs (subcutaneously) and sheep (intravenously). The specific paretic syndrome, krimpsiekte , on the other hand, was reproduced only by the repeated intravenous administration of smaller doses of the 2 unknown bufadienolides to sheep. Histopathological examination revealed a mild to severe multifocal cardiomyopathy in sheep receiving plant material or bufadienolides .

Isolation and characterization of and some observations on poisoning by bufadienolides from Cotyledon orbiculata L. var. orbiculata.

The approximate LD50 and cumulative effect of 4 bufadienolides, isolated from Cotyledon orbiculata, were determined in guinea-pigs. Two of the bufadienolides proved to be mildly cumulative. One of the mildly cumulative bufadienolides induced subacute intoxication with some signs of krimpsiekte when injected intravenously into a sheep over 13 days. In the sheep, the paralysis was accompanied by signs of malaise, such as electrocardiographic changes and ruminal stasis.