A 90-day feeding study of the alkaloids of Lupinus angustifolius in the rat.

Groups of 20 Sprague-Dawley rats of each sex were fed diets containing lupin alkaloid at dose levels of 0, 100, 330, 1000 and 5000 ppm supplemented with maltodextrin to attain a level of 4.5%, for 13 wk (equivalent to average daily intakes of lupin alkaloid of approximately 0, 10, 30, 100 and 500 mg/kg body weight/day, respectively, over the course of the study). A further group of rats was fed control (basal) diet over the same period. All control and high-dose animals underwent an ophthalmological examination before the start of the study and before autopsy. Blood samples were collected from all rats prior to the start of treatment, during wk 6 and prior to autopsy for haematological and clinical chemistry examination. All animals were monitored daily for change in clinical condition, and body weight and food intake were measured twice weekly. A range of tissues were preserved for histological examination at autopsy. There was an initial drop in food intake by all rats in the 1000 and 5000 ppm groups and thereafter the intake was between 90% and 95% of that of the controls. In general, no other effects related to treatment were seen. On the basis of the lower body weights and food intakes of the groups fed the alkaloid at levels of 1000 and 5000 ppm, a no-observed-adverse-effect level (NOAEL) of 330 ppm is seen under the conditions of this study. It is likely that these effects are entirely due to the antipalatability effect of the lupin alkaloids. In view of the growth rates, haematology, clinical chemistry and histological findings, a speculative NOAEL of 1000 ppm may be more appropriate.

Contribution of cobalamin analogues to plasma vitamin B12 concentrations in cattle.

Plasma vitamin B12 concentrations in cattle were analysed by a radioisotope dilution assay using pig intrinsic factor and a microbiological assay using Euglena gracilis. Both assays provided similar results for samples of cattle plasma containing vitamin B12 concentrations ranging from 0.07 to 3.60 micrograms litre-1 (r = 0.95, P less than 0.001). The addition of excess cobinamide in the radioisotope dilution assay to block non-specific binding in the intrinsic factor preparation due to the presence of R-type binders, was used to determine the presence of cobalamin analogues. Cobalamin analogues accounted for up to 50 per cent of the total vitamin B12 concentration in samples of plasma from cows but were virtually undetectable in plasma from sheep.

Absorption by sheep of dieldrin from contaminated soil.

OBJECTIVE: To study the accumulation of dieldrin residues in sheep from ingestion of contaminated soils was studied in two experiments. DESIGN: A controlled feeding study of sheep fed contaminated soils of different type at varying intervals. ANIMALS AND PROCEDURE: Thirty-four 2-year-old wethers were divided into four groups (one control sheep only) and fed water-soluble dieldrin or soil contaminated with aldrin and dieldrin at varying intervals in the first study. In a second study 34 similar sheep were divided into four treatments with one being a control. Sheep were fed sandy, high clay or high organic matter soils with similar dieldrin and aldrin concentrations. RESULTS: In the first study the concentration of dieldrin in the body fat of sheep dosed with dieldrin-contaminated soil was about half that in the body fat of sheep dosed with an equivalent amount of water-soluble dieldrin. The concentration of dieldrin was almost the same in sheep fed 500 micrograms of total dieldrin per day as it was in sheep fed 5000 micrograms every tenth day, over a 50-day period. In the second experiment sheep accumulated nearly three times as much pesticide from a soil with a high organic matter content, and about four times as much from a soil with a high clay content, as from a sandy soil with the same dieldrin content, over a 100-day period. The half-life of dieldrin in the fat of all sheep varied between 96 and 116 days after sheep caused ingesting contaminated soil. CONCLUSIONS: Dieldrin concentrations in the fat of sheep that consume dieldrin contaminated soil fall within 10 days of removal from the source of contamination. However, dieldrin accumulates in the wool of sheep that consume dieldrin contaminated soil.

Treatment of ovine annual ryegrass toxicity with chlordiazepoxide.

The results of pen trials indicate that sheep affected by annual ryegrass toxicity can be treated successfully with chlordiazepoxide. In the first experiment, 87% of chlordiazepoxide-treated animals survived a lethal dose of toxic ryegrass. In the second experiment, 90% of chlordiazepoxide treated animals survived compared with a 10% recovery of control animals.

Soil and agronomic factors associated with cadmium accumulations in kidneys of grazing sheep.

Mean concentration of cadmium (Cd) in kidneys of hogget sheep from 67 flocks grazing in the Agricultural Region of Western Australia was tested for association with soil, pastoral, climatic and nutritional factors. Hoggets grazing pastures on acidic soils and soils with a sandy-textured surface had higher Cd concentrations in kidneys than hoggets grazing pastures on more alkaline soils or soils with a clay-textured surface. Application of more than 100 kg of phosphatic fertiliser during the past 3 years to loamy soils was also associated with greater Cd concentration in kidneys of the grazing animals.

Control of ovine lupinosis: experiments on the use of fungicides.

Experiments were conducted in 4 successive years to assess whether certain fungicides when applied to lupins would control the disease, ovine lupinosis. Results were variable, ranging from failure of fungicidal sprays to prevent the disease in 1969/70 to partial success in 1970/71. Fumigation with methyl bromide was very effective. It is concluded that none of the fungicides tried could be considered as a practical means of controlling the disease.

Residues in beef cattle accidentally exposed to commercial heptachlor.

The changes in concentration of heptachlor epoxide (HCE) and oxychlordane (OCD) were studied in the tissues of beef cattle that had previously grazed pasture contaminated with commercial heptachlor. In 25 cows and heifers monitored over 488 days, the mean concentration of HCE decreased from 22.0 to 0.08 mg/kg and of OCD from 5.68 to 0.18 mg/kg. The respective half-lives were estimated to be 66 and 92 days. In a controlled feeding experiment, nine steers were subjected to one of three dietary regimes; a high plane of nutrition for 81 days then a low plane for 140 days; a moderate plane of nutrition for the total period; and a low plane of nutrition for 81 days then a high plane for 140 days. Approximately five months later similar treatments were applied for 101 and 94 days. The rate of decline of HCE and OCD was consistently greatest in animals on the high plane of nutrition and lowest in those on the low plane. Lactation did not appear to have a major effect on the rate of decline in the tissue concentrations of these compounds. No significant (P greater than 0.05) relationships were found between the concentrations of HCE or OCD in the subcutaneous fat and either milk or blood at any stage of the experiment. At slaughter, from 18 to 24 months after the commencement of the experiments, significant correlations (P less than 0.05) were found among concentrations of HCE and OCD in subcutaneous fat, renal fat and fat from the bone marrow.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of selenium supplementation on immunity, and the establishment of an experimental Haemonchus contortus infection, in weaner merino sheep fed a low selenium diet.

Immunity in 12 weaner Merino sheep fed a low selenium (Se) diet (low Se sheep) was compared with that in 10 matching sheep fed the same diet but each given an intraruminal Se pellet (high Se sheep), while the sheep were housed in individual, sheltered pens. All sheep were challenged with killed Brucella abortus cells (days 0 and 28), rabbit red blood cells (days 0, 7 and 28) and corynebacterium pseudotuberculosis toxoid (days 0 and 28), and serum antibody titres were measured weekly for 8 weeks from day 0. The sheep were then experimentally infected with Haemonchus contortus, and slaughtered 8 weeks later. The mean antibody titre to B. abortus, measured by 4 different tests, was significantly higher in the high Se sheep on occasions during the primary immune response phase (Rose Bengal test - day 21 (p less than 0.05), day 28 (p less than 0.025); complement fixation - day 7 (p less than 0.05); enzyme-linked immunosorbent assay - day 14 (p less than 0.01); serum agglutination - no differences), but not during the secondary phase. The mean antibody titre to rabbit red blood cells, measured by haemagglutination test, was marginally higher in the high Se sheep on day 49 (p = 0.049). The mean antibody titre to C. pseudotuberculosis, measured by enzyme-linked immunosorbent assay, was not significantly different between the groups at any time during the trial. In addition, the mean in-vitro responsiveness of peripheral blood lymphocytes to stimulation with phytohaemagglutinin in the high Se sheep was significantly greater than that in 10 sheep from the low Se group on day 22 (p less than 0.01), but not day 50.(ABSTRACT TRUNCATED AT 250 WORDS)

Control of alpha-mannosidosis in Angus cattle.

Tests for the detection of animals heterozygous for alpha-mannosidosis were undertaken on samples taken from 34,203 cattle registered with the Angus Society of Australia. Results indicates 1,836 (5.4%) of the animals were heterozygotes. Heterozygotes were detected in 214 (51%) of the herds examined.

Erythrocyte selenium-75 uptake as a measure of selenium status in weaner sheep, and its relationship to erythrocyte glutathione peroxidase activity.

The relationship between in vitro erythrocyte 75Se uptake (75Se uptake) and erythrocyte glutathione peroxidase (EGSHPx) activity was examined in weaner sheep during periods of selenium depletion and repletion, to determine whether 75Se uptake was better correlated than EGSHPx activity to the development of weaner nutritional myopathy. In the 2 trials conducted, only 3 of 45 Merino wether weaners developed clinical myopathy and histological lesions in skeletal muscles. The 75Se uptake values and EGSHPx activities in these 3 sheep were no different from those in the unaffected sheep. There was a significant negative correlation between 75Se uptake values and EGSHPx activities over the entire period of the trials. It could not be demonstrated that 75Se uptake was any better correlated than EGSHPx activity to the development of nutritional myopathy, and it was concluded that EGSHPx activity indicated selenium status better than 75Se uptake in weaner sheep.

The pathogenesis of lupinosis in the rat.

The morphological, histochemical and biochemical changes in the liver of rats challenged with a single intraperitoneal injection of the toxin of lupinosis were investigated. The resulting injury is biphasic. The initial non-specific phase lasts for a few days, affects mainly the centrilobular region and is characterised by mitochondrial swelling, lipid accummulation and autophagosome formation. In addition, the hepatic concentrations of OCT, LDH, SDH, and AST fall while that for ALP increase. This is accompanied by a rise in the serum concentration of many of these enzymes. The second phase lasts longer and is characterised by phenomena indicative of abnormalities in hepatocytic division. Normal and abnormal metaphase as well as multinucleated hepatocytes falls. The possibility that the toxin of lupinosis interferes with microtubular units is considered.

Absorption of zinc from lupin (Lupinus angustifolius)-based foods.

The absorption of Zn from a lupin (Lupinus angustifolius) milk fortified with Ca, a bread containing lupin flour (230 g/kg), a sauce containing lupin flour and a sauce containing a lupin-protein isolate was determined in humans by measuring the whole-body retention of radioisotope from meals labelled with 0.02 MBq 65Zn, allowing for endogenous excretion of Zn, after 14 d. The absorption of Zn from the Ca-enriched milk (16.2%) and the bread made with lupin flour (27.0%) was similar to literature figures for comparable soya-bean products. The absorption from composite meals made with lupin flour (28.2%) and protein isolate (32.7%) was significantly higher than that reported for comparable soya-bean products. In a second experiment the absorption of Zn from a lupin-milk base and a soya-bean-milk base was compared with that from Ca-supplemented bases. The absorption of Zn from the lupin-milk base (26.3%) was significantly higher than from the soya-bean-milk base (17.6%), and neither was significantly altered by the addition of Ca. Overall the absorption of Zn from lupin-protein foods was found to be higher than from comparable soya-bean products. Lupin milk could be an attractive alternative to soya-bean milk for infant formulas.

TREATMENT OF OVINE ANNUAL RYEGRASS TOXICITY WITH CHLORDIAZEPOXIDE.

The results of pen trials indicate that sheep affected by annual ryegrass toxicity can be treated successfully with chlordiazepoxide. In the first experiment, 87% of chlordiazepoxide-treated animals survived a lethal dose of toxic ryegrass. In the second experiment, 90% of chlordiazepoxide treated animals survived compared with a 10% recovery of control animals.

A comparison of two bioassays used to estimate the toxicity of lupin material infected with Phomopsis leptostromiformis.

Eighty-four lupin samples were tested for toxicity using two currently available bioassays. One utilizes sheep and the other nursling rats. There was an association between the results obtained by the two bioassays but 65% of the variance of the nursling rat bioassay results was not associated with the sheep bioassay results in the linear regression. Reasons for this poor correlation are suggested and discussed.

Acute toxicity of the major alkaloids of cultivated Lupinus angustifolius seed to rats.

The seed of modern cultivars of Lupinus angustifolius normally contain less than 0.03% alkaloids. The acute oral LD50 to rats of a pro rata mixture of the alkaloids of L. angustifolius seed was found to be 2279 mg/kg. For lupanine the LD50 by oral administration was 1464 mg/kg and by intraperitoneal injection 177 mg/kg. For 13-hydroxylupanine the LD50 by intraperitoneal injection was 199 mg/kg. Since these two alkaloids comprise about 85% of the total and are known to be rapidly cleared from the body it is suggested that the alkaloids in this species do not pose a health problem for man.