Molasses as a possible cause of an "endocrine disruptive syndrome" in calves.

During the mid 1990s a potentially serious, chronic syndrome was reported in well-managed beef and dairy herds from unrelated parts of South Africa. Farmers reported that it manifested as various combinations of decreased production, decreased weaning masses, apparent immune breakdown in previously immunocompetent animals, increased reproductive disorders, various mineral imbalances in non-deficient areas and goitre, noticeable as enlarged thyroid glands. The farmers associated this syndrome with certain batches of sugar cane molasses and molasses-based products. The syndrome was reminiscent of an "endocrine disruptive syndrome". The objective of this study was to evaluate the suspected endocrine disruptive effect of molasses included in cattle feed. Using existing in vitro assays, four batches of molasses syrup were screened for possible inclusion in a calf feeding trial. Two batches were selected for the trial. Thirty-two, 4- to 6-week-old, weaned Holstein bull calves were included in the single phase, three treatment, parallel design experiment. In two of the groups of calves, two different batches of molasses were included in their rations respectively. The control group was fed a ration to which no molasses was added, but which was balanced for energy and mineral content. The mass gain of the calves was recorded over the 6-month study period. The calves were clinically examined every week and clinical pathology parameters, immune responses and endocrine effects were regularly evaluated. Even though endocrine disrupting effects were detected with the in vitro screening assays, these could not be reproduced in the calves in the experiment. The two batches of molasses utilized in the calf feeding trial did not induce major differences in any of the parameters measured, with the exception of a lower mass gain in one of the molasses-fed groups (Group 1), which tended towards significance. The results of the study indicate that the two batches of molasses had no endocrine disruptive or immunosuppressive effects in calves.

Confirmed Datura poisoning in a horse most probably due to D. ferox in contaminated tef hay.

Two out of a group of 23 mares exposed to tef hay contaminated with Datura ferox (and possibly D. stramonium) developed colic. The 1st animal was unresponsive to conservative treatment, underwent surgery for severe intestinal atony and had to be euthanased. The 2nd was less seriously affected, responded well to analgesics and made an uneventful recovery. This horse exhibited marked mydriasis on the first 2 days of being poisoned and showed protracted, milder mydriasis for a further 7 days. Scopolamine was chemically confirmed in urine from this horse for 3 days following the colic attack, while atropine could just be detected for 2 days. Scopolamine was also the main tropane alkaloid found in the contaminating plant material, confirming that this had most probably been a case of D. ferox poisoning. Although Datura intoxication of horses from contaminated hay was suspected previously, this is the 1st case where the intoxication could be confirmed by urine analysis for tropane alkaloids. Extraction and detection methods for atropine and scopolamine in urine are described employing enzymatic hydrolysis followed by liquid-liquid extraction and liquid chromatography tandem mass spectrometry (LC/MS/MS).

Claviceps cyperi, a new cause of severe ergotism in dairy cattle consuming maize silage and teff hay contaminated with ergotised Cyperus esculentus (nut sedge) on the Highveld of South Africa.

During December/January 1996/97 typical summer syndrome (hyperthermia and a 30% drop in milk yield) occurred in succession in two Holstein dairy herds (n=240 and n=150 milking cows, respectively) on the South African Highveld. These farms are situated in the midst of the prime maize and dairy farming areas of South Africa where this condition had never been diagnosed before. The individual components of the concentrate on both farms were negative for ergot alkaloids. Endophytic fungi and/or ergot infestation of teff and other grasses fed to the cows were then suspected of being involved, but neither endophytes nor ergot alkaloids could be implicated from these sources. By measuring the serum prolactin levels of groups of sheep (n=5) fed the first farm's total mixed ration (TMR) or its three individual fibre components for a period of 11 days, the source of the ergot alkaloids was identified. A statistically significant decrease in the level of this hormone occurred only in the group on maize silage (which constituted 28% on dry matter base of the TMR). The involvement of the maize silage was further chemically confirmed by the high levels of total ergot alkaloids, predominantly ergocryptine, found by LC-MS in the silage as well as in the TMR (115-975 ppb and 65-300 ppb, respectively). The ergot alkaloid content (mainly ergocryptine) of the maize silage on the second affected farm was 875 ppb. Withdrawal of contaminated silage resulted in gradual recovery of stock on both farms. Nut sedge (Cyperus esculentus and Cyperus rotundus of the family Cyperaceae) has a world-wide distribution and is a common weed in annual crops, and can be parasitized by Claviceps cyperi. Careful examination of the maize silage from both farms revealed that it was heavily contaminated with nut sedge and that it contained minute sclerotia, identified as those of Claviceps cyperi, originating from the latter. Nut sedge was abundant on both farms and it is believed that late seasonal rain had resulted in mature, heavily ergotised nut sedge being cut with the silage. Claviceps cyperi sclerotia, collected on the affected fields in the following autumn contained 3600-4000 ppm ergocryptine. That the dominant alkaloid produced by this particular fungus was indeed ergocryptine, was confirmed by negative ion chemical ionization MS/MS. In one further outbreak in another Holstein herd, teff hay contaminated with ergotised nut sedge and containing 1200 ppb alkaloids, was incriminated as the cause of the condition. This is the first report of bovine ergotism not associated with the Poaceae infected with Claviceps purpureum or endophytes but with the family Cyperaceae and this particular fungal phytopathogen.

Datura contamination of hay as the suspected cause of an extensive outbreak of impaction colic in horses.

Datura poisoning of horses is extensively reviewed. An outbreak of intractable impaction colic affecting 18 of 83 horses was stopped by withdrawing dried tef hay contaminated with young Datura plants. The dried, botanically identified Datura stramonium and D. ferox contained respectively 0.15% mass/mass (m/m) hyoscyamine as well as 0.16% m/m hyoscine (scopolamine) and only hyoscine at a concentration of 0.11% m/m. Immature, unidentifiable plants resembling D. stramonium, contained 0.14% m/m and 0.12% m/m of the 2 respective tropane alkaloids. The outbreak was characterised by protracted and repeated colic attacks due to impaction of the large colon and/or caecum without any other anti-muscarinic signs. Comparative analyses of single specimens of dried seed of the 2 species collected from both fertilised and waste areas revealed that young South African Datura spp. had levels of tropane alkaloids comparable to those in the well-known toxic seed and were, consequently, equally toxic. The inherent danger of tef hay being contaminated with Datura is emphasised. To our knowledge this is the 1st field case of poisoning in horses ascribed to the vegetative parts of Datura spp.

A lysosomal storage disease induced by Ipomoea carnea in goats in Mozambique.

A novel plant-induced lysosomal storage disease was observed in goats from a village in Mozambique. Affected animals were ataxic, with head tremors and nystagmus. Because of a lack of suitable feed, the animals consumed an exotic hedge plant growing in the village that was identified as Ipomoea carnea (shrubby morning glory, Convolvulaceae). The toxicosis was reproduced by feeding I. carnea plant material to goats. In acute cases, histologic changes in the brain and spinal cord comprised widespread cytoplasmic vacuolation of neurons and glial cells in association with axonal spheroid formation. Ultrastructurally, cytoplasmic storage vacuoles in neurons were membrane bound and consistent with lysosomes. Cytoplasmic vacuolation was also found in neurons in the submucosal and mesenteric plexuses in the small intestine, in renal tubular epithelial cells, and in macrophage-phagocytic cells in the spleen and lymph nodes in acute cases. Residual alterations in the brain in chronic cases revealed predominantly cerebellar lesions characterized by loss of Purkinje neurons and gliosis of the Purkinje cell layer. Analysis of I. carnea plant material by gas chromatography-mass spectrometry established the presence of the mannosidase inhibitor swainsonine and 2 glycosidase inhibitors, calystegine B2 and calystegine C1, consistent with a plant-induced alpha-mannosidosis in the goats. The described storage disorder is analogous to the lysosomal storage diseases induced by ingestion of locoweeds (Astragalus and Oxytropis) and poison peas (Swainsona).

A high-performance liquid chromatographic method for the determination of monofluoroacetate.

A simple isocratic high-performance liquid chromatographic (HPLC) method for the quantitative analysis of monofluoroacetic acid (MFA), the toxic substance of Dichapetalum cymosum, in plant material, rumen contents (gastric contents), and liver samples is described. A suitable HPLC column that gives optimum sensitivity, accuracy, precision, and separation of MFA is identified. A C-610 organic acid analysis column at ambient temperature with 0.02M H3PO4 as an eluent and ultraviolet detection at 210 nm is utilized to quantitate MFA. Using this method, the average percentage recovery in plant material, bovine liver, and rumen samples is 94.8%, and a detection limit of 12 microg/L is achievable.

Studies on South African cardiac glycosides. II. Observations on the clinical and haemodynamic effects of cotyledoside.

In guinea-pigs, the oral and subcutaneous LD50 values were very similar (cf. 0,173 mg/kg over 48 h with 0,116 mg/kg over 24 and 48 h). When dosed subcutaneously, a cumulative effect was observed. Intravenous administration of cotyledoside to anaesthetized guinea-pigs resulted in: dyspnoea, increased heart rates and blood pressures, and electrocardiagraphic changes typical of cardiac glycoside poisoning. A positive cardiac inotropic effect was succeeded by a positive chronotropic one. In sheep, acute and subacute intoxication resulted in ruminal, respiratory and cardiac changes. The signs included ruminal stasis, cyanosis, cardiac arrhythmia, ectopic foci and AV dissociation, followed by hypotension and progressive respiratory and cardiac failure. The skeletal muscles were affected in only 1 sheep vide infra. In chronically intoxicated sheep typical clinical signs of "krimpsiekte" developed, e.g. weakness, reluctance to stand, unsteadiness on feet, tremor and paresis of hindquarter muscles, paresis of the neck, arching of the back and standing with the feet close together. Respiratory function was affected in all 3 cases; ruminal stasis, with concomitant loss of appetite occurring in one, and a transient change in heart function in another. The syndrome induced by acute cotyledoside poisoning is similar to that of other cardiac glycosides, but the paretic signs of chronic intoxication resemble "krimpsiekte", a disease associated only with intoxication with the plants of the family Crassulaceae.

The distribution, diagnoses and estimated economic impact of plant poisonings and mycotoxicoses in South Africa.

Maps are provided showing the national and provincial distributions of many plant poisonings and mycotoxicoses in South Africa. The various poisonings are briefly described and criteria for their diagnoses are given. The annual mortalities of cattle from plant poisonings/mycotoxicoses in South Africa were calculated as c. 37 665 head with a current cash value of c. R57 627 450, and that of small stock as c. 264851 head with a value of c. R46 878 627. The annual total cost of plant poisonings/mycotoxicoses to the livestock industry of South Africa is conservatively estimated at R104 506 077. These figures do not include hidden losses such as diminished production, reproductive failure, the cost of not utilizing toxic pastures and the fall in price of infested land.

Furunculosis in rainbow trout (Salmo gairdneri) raised in sea water.

Ulcerative skin lesions were encountered in rainbow trout raised in sea water by a commercial concern in the Western Cape, South Africa. Grossly, the lesions resembled furunculosis but, histopathologically, they differed from typical furunculosis in that bacterial colonies were rarely found in the organs, and also the kidneys and spleens were minimally involved. The causative organism was identified as an achromogenic Aeromonas salmonicida that shared characteristics with all 3 subspecies, salmonicida, masoucida and achromogenes. This is the first report of an outbreak of this disease in South Africa.

Leukoencephalomalacia: a mycotoxicosis of Equidae caused by Fusarium moniliforme Sheldon.

When 2 horses were dosed with cultures of a Fusarium moniliforme isolate that had previously caused only hepatosis, 1 developed brain oedema and hepatosis, and the other only leukoencephalomalacia. A 3rd horse developed both leukoencephalomalacia and hepatosis after being dosed with another isolate obtained from maize which was associated with a natural outbreak of the nervous form of the disease. Since leukoencephalomalacia and hepatosis could be induced by the same culture material, it was concluded that both syndromes were manifestations of the same toxicosis. There was also some evidence that leukoencephalomalacia might be specifically induced by the administration of smaller doses of the culture material to horses over a longer period. The clinical signs of nervous disorder included ataxia, paresis, apathy, hypersensitivity, frenzy, and other locomotory and psychic disturbances. Autopsy showed that the brains were oedematous, and focal areas of liquefactive necrosis were present in the cerebral white matter. In 1 case the malacic areas were not confined to the subcortical white matter but were microscopically visible in the cerebral cortex as well. An histopathological examination of the areas bordering on the malacic areas revealed rarefication of the white matter, perivascular haemorrhages, oedema and cellular infiltration composed mainly of plasma cells and eosinophiles. Many of the macrophages in these areas contained lipfuscin-like granules, but these granules also occurred extracellularly in the neuropil. In the layers of the cortex nearest the malacic areas, satellitosis and neurophagia were commonly seen.

Investigation of biological samples for monofluoroacetate and Dichapetalum cymosum poisoning in southern Africa.

A need has existed for the accurate identification of monofluoroacetate (MFA) poisoning in southern Africa. The development of a new method for the determination of MFA has made the analysis of a variety of biological samples (n = 50) feasible. The method has been used in the laboratory over 24 months. Monofluoroacetate was present in 66% of samples from cases of suspected poisoning, reflecting the extent of the problem. Stability of MFA in samples was also determined so as to have a time-bound baseline for the acceptance of samples submitted. It was found that there was a decrease in the level of MFA and, after 14 days at room temperature, only 50% of the spiked dose could be identified. It is suggested that samples be analyzed within 7 days of mortality if they not kept frozen.

Evaluation of the efficacy of d-penicillamine and trientine as copper chelators using an in vitro technique involving ovine red blood cells.

An in vitro technique for haemolysing ovine red blood cells with copper sulphate was standardized to induce c 50% haemolysis with 0.5 mM CuSO4 after incubation for 14 h at 38 degrees C. This technique was then applied to test the efficacy of trientine and d-penicillamine in preventing haemolysis. Trientine concentrations of 0.5; 1.0 and 1.5 mM were found to be the most effective (P < 0.05) in reducing copper-induced haemolysis. One and 1.5 mM concentrations of d-penicillamine were also effective (P < 0.05), but in this experiment a 0.5 mM concentration failed to protect the erythrocytes.

Observations on the clinical, cardiac and histopathological effects of fluoroacetate in sheep.

Fluoroacetate was dosed per stomach tube to 17 Merino sheep at the rate of 0,05-1,0 mg/kg/day. The clinical signs, haemodynamic changes, chemical pathology and pathology of acute, subacute and chronically intoxicated cases are described. Tetanic convulsions were seen in acutely intoxicated animals and in them respiratory failure, occurring concomitantly with cardiac failure, may have been the cause of death. Subacute intoxication resulted in less conspicuous clinical signs when the sheep were at rest, but they developed apparent nervous signs on being handled, and later tended to lie down. Chronically intoxicated animals were only mildly affected. At all levels of intoxication changes in the chemical pathological parameters were either absent or were mild and transient. The microscopic lesions in the hearts of acutely intoxicated sheep included degeneration as well as necrosis of individual or small groups of myocardial fibres. In the subacutely and chronically intoxicated animals the multifocal myocardial lesions were more widespread and in various stages of development or resolution.

Trout mortalities as a result of Streptococcus infection.

Excessive mortalities were experienced in the bigger rainbow trout (Salmo gairdneri) at an efficiently managed trout farm. All tests for known toxins in the feed and water proved to be negative. A faecal Streptococcus which belongs to the Lancefield group D but which could not be identified as belonging to any of the recognized species within this group, was isolated from the spleens, livers and kidneys of affected fish. Pathogenicity studies with this organism proved it to be highly fatal to trout but not to Mozambique bream (Sarotherodon mossambicus), banded bream (Tilapia sparrmenii), carp (Cyprinus carpio) or largemouth bass (Micropterus salmoides). The isolation and biochemical characteristics of the organism are described. The symptoms, gross- and histopathology of this disease are described and discussed. The disease resembles a haemorrhagic septicaemia and appears to be associated with intensification and conditions of stress.

A highly fatal tremorgenic mycotoxicosis of cattle caused by Aspergillus clavatus.

During February 1975, a tremorgenic neurotoxicosis decimated a herd of cattle in the northern Transvaal. This hitherto unidentified disease was characterized by hypersensitivity, incoordination, a peculiar stiff-legged gait of the hind legs, severe generalized tumors of the sketetal muscles, progressive paresis, paralysis and constipation. The most notable gross pathological lesions were degenerative and necrotic changes in certain skeletal muscles, haemorrhages on the serosal surfaces, especially on the dorsal aspect of the rumen, and gastro-intestinal stasis. Microscopical examination of the central nervous system revealed cytopathological changes consisting of degeneration and necrosis of the large motor cells in the ventral horns of the spinal cord and bigger neurones in numerous nuclei of the medulla oblongata, midbrain and thalamus. By feeding the suspect ration and its component parts to cattle and sheep, it was possible to identify mouldy sorghum beer residue as the toxic component in the ration. A. clavatus, the dominant fungus on the toxic residue, was readily isolated in pure culture. The entire syndrome was then reproduced in a yearling Friesland steer dosed with pure cultures of the A. clavatus isolate grown on autoclaved non-toxic sorghum beer residue. The toxic principle is not known, but it does not appear to be patulin, tryptoquivalone, tryptoquivaline, or any other known tremorgen.

Experimental studies with Stronglyloides papillosus in goats.

Unusual clinical and pathological observations in the field in goats and sheep suffering from Strongyloides papillosus infection prompted experimental work on this parasite. Goats were infected percutaneously with either single or multiple, low or high levels of S. papillosus. Young goats up to 12 months of age were found to be the most susceptible. Some animals, however, showed substantial resistance to infective doses. Clinical signs included transient diarrhoea, misshapen, elongated faecal pellets terminally, dehydration, anorexia, cachexia, gnashing of teeth, foaming at the mouth, anaemia and nervous signs such as ataxia, a wide-based stance, stupor and nystagmus. A 'pushing syndrome' was seen in 22% of the animals. The pathological changes are described and included enteritis, status spongiosus in the brain, hepatosis leading to rupture of the liver, nephrosis, pulmonary oedema, interstitial pneumonia and pneumonia. About 6% of the goats died acutely from fatal hepatic rupture. The development of an acquired immunity was determined. The immunity elicited an allergic skin reaction at the application site of larvae or injection sites of larval metabolites. This immunity, however, could be breached by large doses of larvae. The most profound clinicopathological changes induced by the parasites were an anaemia (most pronounced in the young goats) and hypophosphataemia. Trace element analyses provided evidence of Cu, Mn and possibly Se deficiencies in some goats.

Plant poisonings and mycotoxicoses of importance in horses in southern Africa.

Well-known plant poisonings such as 'dunsiekte' (seneciosis) and 'jaagsiekte' (crotalariosis) of horses in southern Africa are briefly reviewed. Relatively unfamiliar mycotoxicoses such as stachybotryotoxicosis and perennial rye grass staggers and potentially occurring exotic intoxications such as equine nigropallidal encephalomalacia and ergot alkaloid poisoning are also discussed. This article is aimed at informing the southern African equine practitioner about probable poisonings that might occur locally in horses.

Suspected poisoning of puppies by the mushroom Amanita pantherina.

Amanita pantherina poisoning is suspected as the cause of a severe, transient neurological disorder in three 5-week-old German shepherd puppies. There was very strong circumstantial evidence that this mushroom had been eaten, and the signs encountered were similar to those described in confirmed field cases of intoxication in dogs. It was also in many respects consistent with the syndrome seen in humans. A veterinary perspective on the hallucinogenic Amanita spp. is given and the veterinary literature on mushroom intoxication is briefly reviewed as, in contrast to humans, it is not often reported in animals. This is the 1st report of suspected mushroom intoxication of animals in South Africa.

Mortality in chickens associated with blister beetle consumption.

Mortality in young chickens was associated with blister beetle consumption. Two species of these insects, Cyaneolytta sp. and Cylindrothorax sp., were found in the chickens' crops, and erosive lesions in the gastrointestinal tract were compatible with blister beetle poisoning (cantharidiasis).

Bothriocephalosis in the common carp in the Eastern Transvaal.

The first outbreak of bothriocephalosis in common carp in South Africa is recorded. This condition was caused by Bothriocephalus acheilognathi, a pseudophyllid tapeworm not previously identified in this country. Methods to limit its spread are suggested.