Identification of the glycosidase inhibitors swainsonine and calystegine B2 in Weir vine (Ipomoea sp. Q6 [aff. calobra]) and correlation with toxicity.

The polyhydroxy alkaloid glycosidase inhibitors swainsonine [1] and calystegine B2 [6] have been identified as constituents of the seeds of the Australian plant Ipomoea sp. Q6 [aff. calobra] (Weir vine) by gas chromatography-mass spectrometry and by their biological activity as inhibitors of specific glycosidases. This plant, which is known only from a small area of southern Queensland, has been reported to produce a neurological disorder when consumed by livestock. The extract of the seeds showed inhibition of alpha-mannosidase, beta-glucosidase, and alpha-galactosidase, consistent with the presence of 1 and alkaloids of the calystegine class. Histological examination of brain tissue from field cases of sheep and cattle poisoned by Weir vine showed lesions similar to those observed in animals poisoned by the swainsonine-containing poison peas (Swainsona spp.) of Australia and locoweeds (Astragalus and Oxytropis spp.) of North America. These results indicate that Weir vine poisoning is an additional manifestation of the induced lysosomal storage disease, mannosidosis, possibly exacerbated by inhibition of the enzymes beta-glucosidase and alpha-galactosidase by calystegine B2. This is the first reported example of a single plant species capable of producing structurally distinct glycosidase inhibitors, namely, alkaloids of the indolizidine and nortropane classes.

Tropical pasture hay utilization with slaframine and cottonseed meal: ruminal characteristics and digesta passage in wethers.

Sixteen mature, ruminally cannulated wethers (average BW = 41 +/- 1 kg) were fed a low-quality hay diet with or without a cottonseed meal (CSM) supplement and the parasympathomimetic agonist slaframine (SF). Treatments were basal diet (Mitchell grass hay, 4.8% CP, 46.8% ADF) available on an ad libitum basis, basal diet plus SF (8 micrograms/kg BW, 2 x daily i.m. injection), basal diet plus CSM (41.0% CP; 100 g/d), or basal diet plus SF and CSM. Treatments were arranged as a 2 x 2 factorial within a replicated 4 x 4 Latin square with 20-d periods followed by a 10-d adjustment during which only the basal diet was fed. All measurements were performed within the final 10 d of each period. Slaframine increased salivary flow by 10 to 35% (P < .07), ruminal fluid dilution rate by 8 to 11% (P < .10), and pH by 3 to 4% (P < .001). A twofold increase (P < .05) in ruminal cellulolytic bacteria numbers occurred in SF-treated wethers. Despite these SF-induced changes in the ruminal environment, whole-tract apparent nutrient digestibility, N and mineral balance, and ruminal VFA concentrations were not changed. Cottonseed meal increased forage intake by 34 to 54% (P < .001) and DM digestibility by 30% (P < .001). Cottonseed meal supplementation of a Mitchell grass hay diet improved nutritional status and attenuated live weight loss.

Acquired resistance of sheep to larvae of Lucilia cuprina, assessed in vivo and in vitro.

The effect on subsequent larval survival of infesting sheep repeatedly with larvae of Lucilia cuprina was assayed in vivo and in vitro. One in vivo assay technique, in which implanted larvae were grown to third instar, indicated a significant reduction in larval survival; another in vivo technique, in which larvae were allowed to develop to second instar in small aluminium rings attached to the sheep, indicated no reduction in larval growth or survival. Larvae of Lucilia cuprina grown in vitro on media containing sera from previously infested sheep were significantly retarded in growth after 20 h compared with controls; no difference was detected when larvae were allowed to develop to pupation on two changes of the same media. No significant differences in survival of larvae either to 20 h or to pupation were obtained between the two treatments. ELISA antibody levels against crude soluble larval material were significantly higher for sera from infested sheep than for control sera, and the regression of antibody level on mean larval weight obtained after 20 h growth in vitro was significant. The immunoglobulin fraction isolated from sera of infested sheep significantly retarded larval growth when incorporated with normal serum in growth media. These results are consistent with an effect of specific anti-larval antibody produced by sheep in response to infestation.

Generalised glycogenosis in Brahman cattle.

Generalised glycogenosis was diagnosed in Brahman cattle on 4 Queensland properties on the basis of clinical observations and pathological and biochemical findings. The disease presented as a problem of ill-thrift and poor growth rate in calves which eventually showed nervous signs. Histologically there was vacuolation in the cells of the central nervous system, heart and muscular tissues. Biochemical assay of liver and blood mononuclear cells demonstrated a deficiency of alpha-glucosidase. Parents of affected calves had approximately half the alpha-glucosidase activity of that found in normal cattle.

Acute nicotinamide deficiency in the pig induced by 6-aminonicotinamide.

Pigs given intraperitoneal injections of 6-aminonicotinamide developed a syndrome similar to a natural disease found in Queensland. Focal lesions, involving mainly destruction of glial cells, developed in the intermediate grey matter of the cervical and lumbar enlargement of the spinal cord. There was widespread vacuolation of spinal cord white matter. The glial cell loss was repaired. We suggest that natural acute nicotinamide deficiency can occur if a threshold dose of an antimetabolite of nicotinamide is absorbed over a short period of time.

Clinical studies on tremorgenic mycotoxicoses in sheep.

The clinical responses of sheep dosed orally over seven to 14 weeks with the dried mycelium of a soil-borne mould containing the tremorgenic mycotoxin penitrem A are described. An initial tremoring response was gradually overshadowed by an incoordination syndrome similar to that seen in ryegrass staggers. In spite of protracted staggers symptomatology, correlated with in vitro changes in the release of neurotransmitter amino acids from corpus striatum nerve ending preparations, no histopathology was evident in the wide range of neural and other tissue studied. The biochemical nature of this reversible mycotoxicosis is discussed.

Crofton weed (Eupatorium adenophorum) toxicity in horses.

The clinical signs and pathology of 6 field cases of a respiratory disease of horses which occurs in the coastal hinterland of south-eastern Queensland are described. The condition has occurred for many years and has been thought to have been associated with ingestion of Crofton weed (Eupatorium adenophorum). Coughing, rapid heaving respiration, decreased exercise tolerance and loss of condition were seen in affected horses. In longstanding cases fibrosis, alveolar lining cell proliferation, oedema, neutrophil infiltration and abscessation were seen. In some cases vascular thrombosis and infarction occur in the lungs. Similar signs and lesions occurred in one horse fed E. adenophorum for 8 months and early lesions in another fed the flowering stage of the plant for about 6 weeks. Lesions also developed in 2 rabbits experimentally fed the plant, but not in sheep or rats.

Leucoencephalomyelitis of goat kids.

A leucoencephalomyelitis in 6 goat kids 2 to 5 months old is described. The disease was characterised by fever, ataxia, posterior paresis, circling and hyperaesthesia progressing to prostration. The neural lesion was confined to the white matter of the cerebellum and posterior brain stem in 4 kids, but in 2 others the cervical spinal cord was the main site affected. The lesion was characterised microscopically by dense perivascular cuffing with mononuclear cells, infiltration of the parenchyma with macrophages and a proliferation of glial cells and by a marked primary demyelination. In more advanced lesions, areas of the neurophil were replaced by a loose glial scar. There were associated pulmonary lesions of interstitial pneumonitis and hyperplasia of the peribronchiolar lymphoid tissue. Attempts to isolate an aetiological agent and to transmit the disease to young goat kids and lambs were unsuccessful. The disease has not been reported before in Australia but has distinct similarities to an infectious leucoencephalomyelitis of young goats which has been described in North America.

The identification of Babesia equi in Australia.

A Babesia parasite, isolated from the blood of a horse at Bowral, New South Wales, was identified on the basis of its morphological features, host specificity and serological reactions, as Babesia equi (Laveran 1901). The case was originally reported by Churchill and Best (1976, Aust. vet. J. 52: 487) and is the first record of equine babesiosis in Australia. In preliminary studies, the organism produced only a mild disease in an intact horse, but caused the typical clinical syndrome of acute babesiosis in a splenectomised horse, which died 19 days after the intravenous inoculation of the parasites.

An outbreak of Swainsona poisoning in horses.

On 6 properties in south-western Queensland an outbreak of nervous disease occurred horses due to ingestion of Swainsonia (Darling pea). Loss of condition, depression, hyperaesthesia and hyperexcitability were seen in affected horses. At autopsy of 2 horses generalised c ytoplasmic vacuolation was seen in the neurones of the central nervous system and in the liver, adrenal and thyroid. The clinical and pathological features were similar to those described in horses suffering from Swainsona poisoning in Australia and Astragalus and Oxytropis in North America.

Neurologic lesions in young captive lions associated with vitamin A deficiency.

Ataxia and incoordination occurred in 7 lion cubs in a lion park. Postmortem examination of 3 cubs revealed thickened skull bones which compressed the brain and lead to prolapse of the posterior vermis of the cerebellum through the foramen magnum. In 2 of the lions compression of the anterior cervical spinal cord caused marked damage particularly in the dorsal funiculi. In both cubs the central canal of the spinal cord was abnormal being extended laterally. The pathological changes in the bones and the brain together with low liver vitamin A levels lead to a diagnosis of hypovitaminosis A.

Humpy back of sheep. Clinical and pathological observations.

Humpy back, a disease of Merino sheep in western Queesland, occurs during mustering for shearing. It is usually seen in summer 6-10 weeks after substantial rainfall and is thought to be caused by the ingestion of a toxic plant. The disease is characterised clinically by a short-stepping, stilted gait of the hind limbs, followed by lowering of the head, arching of the back and inability to continue walking. Histopathological examination of 8 cases from 5 properties revealed a Wallerian degeneration of the white matter throughout the length of the spinal cord with the ventral and lateral columns most severely affected. A similar degenerative change was seen in the posterior cerebellar peduncles of 3 of the sheep. A more severe hind limb incoordination with more extensive degeneration of the white matter of the spinal cord, medulla and cerebellum was seen in a case of humpy back of two years duration. Similar, but much milder, spinal cord lesions were found in apparently unaffected sheep from the same group as the sheep affected with humpy back on 2 properties. Severe myodegeneration of hind limb muscle groups was seen in 3 affected sheep. It was thought to be associated with the long rail journey (1500 km) to the laboratory after the sheep were affected in the field.