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).

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.

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.

Gangrenous ergotism in cattle grazing fescue (Festuca elatior L.) in South Africa.

The 1st outbreak of fescue toxicosis in South Africa was recently confirmed in a Brahman herd at Perdekop, near Standerton, in Mpumalanga province, South Africa. Within 3 weeks of being placed on a fescue pasture in mid-winter, 50 of 385 cattle developed lameness and/or necrosis of the tail. The farmer had established Festuca elatior L. (tall fescue, Iewag variety) on c. 140 ha for winter grazing. Fescue may be infected by an endophyte, Neotyphodium coenophialum, which produces ergot alkaloids, in particular ergovaline. Ergovaline concentrations in basal leaf sheaths and grass stems collected during the outbreak ranged from 1720-8170 ppb on a dry-matter basis.

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.

Tremorgenic neuromycotoxicosis in 2 dogs ascribed to the ingestion of penitrem A and possibly roquefortine in rice contaminated with Penicillium crustosum.

Two dogs developed alarming tremorgenic nervous stimulation shortly after ingesting discarded rice that had been forgotten in a refrigerator for an undetermined period and that was covered with a grey-green mould. Both dogs exhibited vomition followed by slight salivation, tremors and ataxia and 1 showed such severe agitation and seizures that it necessitated anaesthesia with thiopentone followed, on recovery, by xylazine. The other dog was only sedated with xylazine. They made an uneventful recovery. The rice vomitus yielded a pure culture of Penicillium crustosum. On chemical analysis it was negative for organochlorine, organophosphor and carbamate insecticides, as well as for strychnine, but contained 2.6 microg/g of the mycotoxins penitrem A as well as 34 microg/g of roquefortine as determined by LC-MS and confirmed by MS-MS. This is the 1st South African case of naturally occurring penitrem A toxicosis and also the 1st case where quantification of the levels of mycotoxins in dog vomitus is reported. The tremorgenicity of roquefortine and its contribution towards this syndrome, is questioned.

Devil's claw (Harpagophytum procumbens) in a Brahman's preputial sheath: a case report from Botswana.

Failure of penile protrusion during attempted service of a cow on heat was investigated in a 3-year-old Brahman bull at Kwakwadi cattle-post in the Kgalahadi sandveld, Kweneng District, Botswana. The investigation revealed that penile protrusion was obstructed by a devil's claw (grapple thorn), a dry fruit of the plant Harpagophytum procumbens, which had lodged in the cavum preputiale. The thorn, which was removed almost completely manually with minimal tissue dissection, had also caused minor lacerations and puncture wounds on the lamina interna pars parietalis. The wounds healed well following treatment with antiseptics and antibiotics and subsequently the bull regained full penile protrusion and served the cows well. This report describes the first case of lodgement of a devil's claw fruit in, and its extraction from, the cavum preputiale of a Brahman.

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.

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.

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).

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.

Noxious toads and frogs of South Africa.

The major defence mechanism in frogs in via the secretion of toxins from their skin. In humans, intoxication may occur when part of the amphibian integument is ingested, as in the form of herbal medicines. Two groups of South African frogs have skin secretions that are potentially lethal to humans and animals. Toads (Bufo and Schismaderma species), the amphibians with which man and his pets most frequently have contact, secrete potent toxins with cardiac glycoside activity. Topical and systemic intoxication, while seen in humans, remains predominantly a veterinary problem. Intoxication by the red-banded rubber frog, which secretes an unidentified cardiotoxin, is far less common. The probable mechanisms of intoxication and management of a poisoned patient are discussed.

Vermeersiekte caused by Geigeria burkei Harv. subsp. burkei var. hirtella merxm. in the northern province of South Africa.

The 1st field outbreak of vermeersiekte induced by Geigeria burkei Harv. subsp. burkei var. hirtella Merxm, is reported. It is also the first recorded outbreak of this disease in the arid sweet bushveld of the Northern Province of South Africa. The toxicosis was experimentally reproduced in a sheep following daily intraruminal administration of 2.5-5.0 g/kg dried, milled plant material for 18 consecutive days. Neither the sheep in the field outbreak nor the ewe in the experiment exhibited any signs of regurgitation of rumen contents (vermeersiekte). All developed only the stiff or paretic/paralytic forms of the disease. Serum activities of CK and GGT were slightly raised in clinically affected sheep (n = 11) during the field outbreak, and serum activities of AST, GLDH, GGT, LDH and CK increased in the ewe dosed with the plant material. Analysis of dried, milled Geigeria plant material confirms that this species is moderately nutritious.

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.

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.