Motor neurone disease in molybdenum-deficient sheep fed the endogenous purine xanthosine: possible mechanism for Tribulus staggers.

BACKGROUND: The occurrence of Tribulus terrestris motor neurone disease (MND) in sheep is linked with grazing Tribulus growing on cultivation paddocks. A previous survey found that the molybdenum (Mo) content of Tribulus growing on uncultivated soils in the Coonabarabran district of New South Wales was 3.03 ppm, but on cultivated soils it was <0.04 ppm. Tribulus contains the purine, xanthosine, which functions as a neuromodulator, and the catabolism of xanthosine is Mo-dependent. DESIGN: To investigate the relationship between xanthosine ingestion and low Mo concentration, eight sheep were fed Mo-deficient lucerne chaff (<0.10 ppm), the Mo antagonist, sodium tungstate, and xanthosine (25 mg/kg/day) over 18 weeks and then returned to pasture. RESULTS: Signs of MND developed in two sheep 30 months later and astrocyte degeneration occurred in all sheep. CONCLUSION: The findings were similar to those observed in sheep with T. terrestris MND, suggesting that the combination of xanthosine ingestion and Mo deficiency may be the cause of this disorder.

Fescue-associated oedema of horses grazing on endophyte-inoculated tall fescue grass (Festuca arundinacea) pastures.

A new form of toxicity called equine fescue oedema is described. The clinical signs included inappetence, depression, and subcutaneous oedema of the head, neck, chest and abdomen. Affected horses had very low plasma albumin values. The toxicity affected 48 of 56 horses on six farms in different states of Australia, and 4 horses have died. All horses were grazing pastures that had been sown with varieties of Mediterranean tall fescue (Festuca arundinacea) that carry the endophyte known as Max P or Max Q. It is proposed that a pyrrolizidine alkaloid, N-acetyl norloline, which is produced by the Max P endophyte, may be responsible for this new toxicity in horses.

Cerebellar ataxia in sheep grazing pastures infested with Romulea rosea (onion grass or Guildford grass).

There have been anecdotal reports since 1962 of 'staggers' in sheep grazing Romulea rosea infested pastures, but this is the first detailed account. In September 2005, a locomotor disorder developed in 12 of 120 Merino wethers that had grazed R. rosea infested pasture at Albury, New South Wales, over several months. Affected sheep displayed signs that included limb paresis, knuckling over in the fetlocks, fine head tremor, incoordination, and an equilibrium disturbance characterised by frequent falling. The microscopic examination of brain and spinal cord tissues from two affected sheep revealed mild vacuolation, occasional lymphocytic cuffing around blood vessels, mild Wallerian degeneration, and occasional glial cells that contained honey-brown cytoplasmic pigments. The most significant changes were found in the cerebellum, where there were decreased numbers of Purkinje cells, increased numbers of glial cells, scattered vacuoles and occasional swollen axons. Previous reports of cerebellar toxicoses in ruminants have involved goats and cattle and have been associated with the ingestion of six Solanum spp. The Purkinje cell loss in this type of disorder is ultimately extensive and consequently affected animals may survive, but will remain permanently disabled.

Abnormal turning behaviour, GABAergic inhibition and the degeneration of astrocytes in ovine Tribulus terrestris motor neuron disease.

OBJECTIVE: To observe the clinical signs of sheep affected by Tribulus terrestris motor neuron disease, to ascertain their response to striatal dopamine reducing drugs, and to examine their brains and spinal cords for microscopic changes. PROCEDURES: Twenty-eight sheep displaying well developed clinical signs of the disorder were observed. Twenty-two of these and 22 normal sheep were then randomly allocated to three groups and treated with diazepam, chlorpromazine, or xylazine. The time that it took an animal to return to a standing position following drug administration was recorded. The brain and complete spinal cord were removed from each of the other six affected sheep and fixed in formalin. Brains were sectioned throughout at 5 mm intervals and spinal cords at 10 mm intervals. All tissues were paraffin embedded and examined by light microscopy. A few samples were examined by electron microscopy. RESULTS: Clinical signs included postural asymmetry with a right:left body-side dominance within the group of 50:50, unequal flaccid paresis in the pelvic limbs, extensor muscle atrophy and adduction of the weaker pelvic limb, and concurrent abduction of the stronger. Forward motion followed either a fixed left or right hand curved trajectory, the sheep no longer being able to choose which. Twelve animals intermittently displayed rotational behaviour that involved loss of postural balance without locomotor activation. The administration of diazepam, chlorpromazine, or xylazine caused limb paresis and sedation, with affected sheep being slower than normal sheep by factors of 8, 3 and 2 respectively, to return to a standing position. There were scattered areas of mild Wallerian degeneration throughout the spinal cord, and in both the brain and the cord there were small numbers of degenerate astrocytes containing novel cytoplasmic pigment granules. CONCLUSIONS: Affected sheep had a dysfunction in the control of directional change and this provides a new insight into the normal mechanism for 'turning' in quadrupeds. Directional change requires a functional asymmetry or lateralisation within the upper motor neuron to accommodate a difference in the rate of forward progression of each body side and, simultaneously, a lateral shift of the centre of gravity. The sensitivity of affected sheep to diazepam is consistent with a pre-existing elevation in GABAergic neuronal inhibition, probably as a result of a reduction in glutamatergic neuronal excitation. The cytoplasmic pigment found in degenerate astrocytes was novel and its presence in the brain nuclei known to contribute to turning behaviour could have aetiological significance. The motor output of the basal ganglia in Tribulus neurotoxicity appeared to be excessively inhibitory to the pelvic limb extensor muscles and was asymmetric, causing fixation of the turning posture but not locomotor activation. An intoxication of specific purine sensitive, glutamate releasing astrocytes, located in nuclei controlling turning, was suspected.

Reassessment of the toxicity of Hypericum perforatum (St John's wort) for cattle.

OBJECTIVE: To investigate the clinical effect of administering sufficient Hypericum perforatum to cattle to deliver quadruple the reported oral toxic dose. ANIMALS: Thirty-six yearling Hereford (n = 18) and Angus (n = 18) steers. DESIGN: A series of six experiments was conducted, each using 12 animals in a 2 x 2 factorial design, with two breeds of cattle (Hereford, Angus) and two dose levels of hypericin, 1.5 mg/kg (treated group) and 0 mg/kg (control group). Each set of 12 steers was used in duplicate experiments, with all animals alternated between treated and control groups. PROCEDURES: Treated groups received finely milled H. perforatum administered orally in gelatin capsules to provide 1.5 mg hypericin/kg body weight. All cattle were then exposed to direct sunlight for 5 h per day for 5 successive days. Rectal temperatures were measured immediately before and at the end of each sunlight exposure session. Rectal temperature above 40 degrees C, together with some other clinical sign of hypericin poisoning, was considered indicative of intoxication. RESULTS: No animals developed a rectal temperature above 40 degrees C or other clinical signs of hypericin poisoning. CONCLUSIONS: While the reported bovine oral toxic dose of 3 g dried plant/kg body weight, for flowering stage, presumed narrow leaved biotype, H. perforatum, is probably correct, the corresponding dose for hypericin of 0.37 mg/kg is incorrect. Based on its known concentration in this plant the toxic dose of hypericin for partially pigmented Hereford-cross cattle is estimated at about 10.5 mg/kg body weight and more than this for fully pigmented cattle. This would imply that cattle of the former type should be about three and a half times better protected against H. perforatum toxicity than are unpigmented, wool protected, Merino sheep. Cattle, particularly if fully pigmented, may have a role in grazing management to control H. perforatum.

The effect of shade, shearing and wool type in the protection of Merino sheep from Hypericum perforatum (St John's wort) poisoning.

OBJECTIVE: To investigate the roles of shade, fleece length and wool type in the protection of sheep from Hypericum perforatum poisoning. ANIMALS: Adult Merino ewes of superfine, fine and medium wool type. DESIGN: Seventy sheep were divided into seven equal groups. During late spring and summer a series of successive, replicate experiments was conducted, each using one group and lasting 5 days. The sheep carried 14 to 24 weeks wool growth. In each experiment the treatments tested were Hypericum +, sunlight + (n = 7); Hypericum +, sun - (n = 1); Hypericum -, sun + (n = 1); Hypericum -, sun - (n = 1). Next, 24 sheep in two equal groups were used in experiments of similar design to the above. Each group consisted of nine recently (1 to 3 weeks previously) shorn and three wool covered (25 to 26 weeks growth) sheep. The treatments tested were Hypericum +, sunlight +, fleece - (n = 9); Hypericum +, sun -, fleece + (n = 1); Hypericum -, sun +, fleece + (n = 1); Hypericum -, sun -, fleece + (n = 1). PROCEDURES: Finely milled Hypericum was administered by gavage to provide 3 mg hypericin / kg body weight. Sheep were sheltered from direct sunlight or were exposed for 5 h per day for 4 successive post-treatment days. Rectal temperatures were measured immediately before and at the end of each sunlight exposure session. Rectal temperature above 40 degrees C was considered indicative of hypericin poisoning. RESULTS: After Hypericum treatment hypericin poisoning was displayed by 26.5% of woolled sheep that were exposed to sunlight, but by none of those that were fully shaded. In similarly treated but recently shorn sheep 94% displayed hypericin poisoning when exposed to sunlight. In the wool covered group the percentages of poisoned animals based on wool type were: superfine 14%, fine 28.5%, medium 33.3%. In the recently shorn group the percentage for all three approached 100%. CONCLUSIONS: A majority of Merinos with at least 14 weeks wool growth will not be poisoned by a single oral dose of 3 mg hypericin /kg, but because hypericin persists in the blood circulation for several days this safe dose will be lowered by continuous daily ingestion. Sheep with access to substantial areas of shade could safely ingest much greater amounts of hypericin. Wool removal greatly increases the risk of poisoning. Superfine Merinos with a wool cover should be able to ingest more hypericin than comparable, medium wool types, without any increased risk of poisoning. The ability of ruminant livestock to safely ingest Hypericum is probably determined more by the amount of skin protection they have against incident sunlight than by differences in hypericin metabolism and excretion capacity.

Evidence that enforced sunlight exposure can cause hyperthermia in cattle ingesting low levels of ergot of rye (Claviceps purpurea), when air temperature and humidity conditions are only moderate.

OBJECTIVE: To determine the rectal temperature response of cattle, following the oral administration of ergot of rye (Claviceps purpurea), under pen conditions of enforced sunlight compared with those of enforced shade. DESIGN AND PROCEDURE: Hereford cross steers were divided into two groups of 18. One group was dosed once, on a Monday morning, with finely ground rye grass ergots at a rate of 180 mg/kg body weight and held in the sun for 7 h each day until Friday afternoon. The other group was not dosed but was similarly held in the sun during the same period. Their rectal temperatures were measured early morning and mid afternoon, from Monday to Friday inclusive. The process was repeated for each group, but this time they were held in the shade. The four treatment options were run concurrently by conducting the experiments over 6 weeks and using 3 animals in each treatment group, each week. RESULTS: The thermic response over all weeks, of the ergot treated, sunlight exposed cattle, was deemed greater than for the other groups, based on the following four parameters. The increase in rectal temperature between early morning and mid afternoon, the size of the mid afternoon rectal temperature rise, the difference between the maximum mid afternoon rectal temperature recorded by an animal in the sun compared with that recorded by the same animal in the shade, and finally the number of animals in a treatment group that recorded rectal temperatures > 40.00 degrees C. The difference in the daily increase in body temperature between the ergot treated, sun exposed cattle and the ergot treated, shaded cattle, was greater than that observed between the sun exposed and shade restricted control cattle. Nine of 18 ergot treated and sun exposed cattle developed hyperthermia; no cattle in the other three groups did. CONCLUSIONS: Some sunlight exposed cattle, dosed with a low amount of ergot of rye, can experience a body temperature elevation above the normal range, even under mild ambient temperature and humidity conditions. Sunlight exposure causes an increase in body temperature in normal cattle that is probably attributable to solar radiant heat. Ergot ingestion causes an increase in body temperature in shaded cattle that is probably attributable to ambient temperature and humidity effects. This response can be increased by sun exposure and this increase is attributable to solar radiant heat effects and possibly also solar radiation. A response to radiant heat is consistent with the known physiological effects of ingested ergot alkaloids, but a response to solar radiation is not.

Seasonal variation in hypericin content of Hypericum perforatum L. (St. John's Wort).

Hypericin and pseudohypericin, bioactive constituents in St. John's Wort (Hypericum perforatum), have been determined in the soft tops of the plant that are most likely to be browsed by foraging livestock. In two consecutive seasons, the hypericin/pseudohypericin concentration in a broad leaf biotype varied from a winter minimum of less than 100 ppm to a summer maximum approaching 3000 ppm. In contrast the narrow leaf biotype increased from similar winter values to summer maxima approaching 5000 ppm. The latter biotype was slower in returning to low levels of hypericin/pseudohypericin.

Sunlight associated hyperthermia as a consistent and rapidly developing clinical sign in sheep intoxicated by St John's wort (Hypericum perforatum).

OBJECTIVE: To assess the usefulness of rectal temperature responses in Australian bred Merino sheep, following the oral administration of Hypericum perforatum (St John's wort), as an early indicator of Hypericum intolerance. DESIGN: Thirty-three Merino ewes were divided into three groups of 11. Each group was dosed with finely ground, dried, flowering growth stage H perforatum plant material at either 5.7, 4.0, or 2.85 g dry plant per kg live weight. This corresponded to 5.3, 3.7 and 2.65 mg hypericin per kg live weight, respectively. PROCEDURE: The sheep were dosed with a plant slurry by stomach tube and then exposed to bright sunlight for up to 5 h per day over successive days. Their clinical responses were observed and rectal temperature measured. RESULTS: Ingestion of H perforatum followed by exposure to bright sunlight frequently resulted in clinical signs attributable to skin irritation and central nervous effects, including an inappropriate increase in body temperature. A decrease in H perforatum ingestion from 5.7 to 2.85 g dry plant per kg live weight and a corresponding decrease in hypericin ingestion from 5.3 to 2.65 mg per kg live weight, was associated with a decrease in the severity of the clinical signs, including the severity of the hyperthermia. CONCLUSIONS: The rectal temperature rise in affected sheep is a reliable indicator of the early development of an adverse clinical effect. There appears to be an absolute requirement for exposure to bright sunlight before any effects of H perforatum will develop. A single dose of H perforatum remains potentially effective for up to 4 days. In the small group of Merino sheep tested a tolerance level for H perforatum, eaten at the flowering stage, of < 1% (plant wet weight) of body weight and a tolerance level for hypericin of < 2.65 mg per kg live weight, were demonstrated.

Chronic gypsum fertiliser ingestion as a significant contributor to a multifactorial cattle mortality.

OBJECTIVE: To assess the validity of claims that heavy metal contamination from an open-cut mine caused the death of 226 cattle on a nearby farm over a period of 18 months, and to investigate other possible contributing factors. PROCEDURE: A retrospective assessment of previous investigations combined with additional chemical analyses. RESULTS: Extensive chemical analyses produced no evidence of heavy metal contamination associated with the mine. Analysis of bones indicated exposure to fluoride in greater than normal amounts. The main source of fluoride seems to have been gypsum that was included in a feed supplement and also ingested from fertiliser dumps on paddocks. The gypsum itself may have contributed significantly to the ill health. Other factors probably affected some classes of animals, notably the young calves. CONCLUSIONS: What originally seemed to be a disease problem of single aetiology probably was an expression of interacting multifactorial causes. This investigation has highlighted the potential toxicity of gypsum to livestock and the need for further studies to establish its basis.