A toxicological investigation of the garden shrub Brunfelsia calcyina var. floribunda (yesterday-today-and-tomorrow) in three species.

Brunfelsia calcyina var. floribunda is an ornamental evergreen shrub found in the United States. A diagnosis of the fatal intoxication of a canine due to consumption of plant material (primarily berries) was made. The significant features of the clinical constellation were similar to those seen with substances interfering with the neurotransmission process, such as lathyrus or strychnine. Necropsy findings on the canine were unrevealing. Toxicologic studies performed on mice and rats with ground shrub material demonstrated that all parts of this plant are toxic, but unequally so. All plant preparations produced signs similar to those of a spinal convulsant. There were no distinguishing gross pathologic or histopathologic findings associated with the toxicoses induced in the laboratory animals with preparations from this plant. The toxic principles from this shrub are water soluble and very stable. The ability of aqueous extracts stored at 4 C to produce the clinical syndrome and subsequent lethality remained unchanged over a period of 4 months. Exposures are not always fatal. They most often occur in the canine and there is a significant hazard for small children.

Case studies on second-generation anticoagulant rodenticide toxicities in nontarget species.

Specimens from 10 cases of second-generation anticoagulant rodenticide poisoning in dogs and cats were submitted to the Texas Veterinary Medical Diagnostic Laboratory during 1986 and 1987. The clinical signs most frequently observed were lethargy, dyspnea, and ventral hematomas; common necropsy findings included hemoperitoneum, hemothorax, and pulmonary hemorrhage. In the instances when histopathological examination of the tissue was done, it supported a diagnosis of coagulopathy. The presence of anticoagulants in serum or liver was confirmed by high pressure liquid chromatography, gas chromatography/mass spectrometry, or a combination of the two. Five cases of brodifacoum poisoning, 2 of bromadiolone, and 3 of diphacinone toxicity were verified. Concentrations of these rodenticides ranged from approximately 0.001 to 12 ppm.

Reproductive effects of fenoxycarb on sheep.

Fenoxycarb (ethyl [2-(4-phenoxyphenoxy)-ethyl] carbamate) is an insect growth regulator used for long-term fire ant control. Because of its effects on insect reproduction and its potential use on pasturage consumed by food animals, a reproductive study was conducted using Rambouillet sheep. The sheep were dosed daily with a placebo or with fenoxycarb at 0.69 or 1.38 mg/kg/day, representing ten (10x) and 20 times (20x) the maximum amounts of fenoxycarb in forage or hay treated at recommended levels for fire ant control. Parameters that were measured included rates of weight gain of adults, serum clinical chemistry profiles of adults, spermatozoal morphology and motility, estrus cycling, pregnancy rates, maintenance of pregnancies to term, numbers of live births, and rates of weight gain of lambs to 28 days. There were no statistically significant (P < or = 0.05) differences between the exposed and control groups of sheep in any of these facets of the study. No clinical signs associated with exposure to fenoxycarb were observed in any animal at any time, and no exposure-related pattern of pathologic lesions or reproductive organ histology was observed. Means of hepatic fenoxycarb residues in the rams followed a statistically significant (P < or = 0.05) dose-related pattern. No fenoxycarb was detected (detection limit of 5 ppb) in any neonatal liver, despite the presence of hepatic fenoxycarb residues in the treated ewes, indicating that transplacental transport of fenoxycarb was minimal. No fenoxycarb was detected in any lamb liver at 28 days, although both the colostrum and the milk of exposed ewes were found to contain fenoxycarb at levels proportional to the treatments. Based on the lack of significant findings in this study, it is unlikely that use of fenoxycarb, according to label instructions (currently applicable to homeowner and registered agricultural usage) for fire ant control in pasturage or hay fields, will affect ruminant reproduction.

Fumonisin B1 concentrations in feeds from 45 confirmed equine leukoencephalomalacia cases.

During the fall of 1989 and winter of 1990, numerous reports of equine leukoencephalomalacia (ELEM) occurred from many regions of the United States. Typically, horses were consuming feed partially or entirely composed of corn and/or corn screenings. From October 1989 through May 1990, samples from 55 confirmed or suspected ELEM cases were received at the National Veterinary Services Laboratories, Ames, Iowa, for fumonisin B1 analysis. Samples from 9 cases in 1984-1985 were also obtained. Fumonisin B1, a mycotoxin produced by Fusarium moniliforme, causes ELEM, but little is known of naturally occurring toxic or safe levels in feeds. To determine what levels of fumonisin B1 in feeds are associated with ELEM, 45 selected cases were studied. The fumonisin B1 concentrations ranged from less than 1 ppm to 126 ppm, with the majority of the samples above 10 ppm. All types of feeds were included: corn, screenings, sweet feeds, and commercially pelleted rations. The length of exposure varied from 7 to greater than 35 days. Horse feed samples not associated with ELEM were also collected and analyzed. None of the nonproblem feed samples contained fumonisin B1 levels greater than 8 ppm.

High pressure liquid chromatographic determination of cantharidin, using a derivatization method in specimens from animals acutely poisoned by ingestion of blister beetles, Epicauta lemniscata.

Experimental animals (rabbit, rat, goat, sheep, and pony) were given cantharidin or dried preparations of blister beetles (Epicauta lemniscata) to stimulate naturally occurring toxicosis in which beetles were ingested with alfalfa hay. A sensitive high-pressure liquid chromatographic method, involving derivatization of cantharidin with p-nitrobenzyloxyamine, was developed to detect the toxin extracts of ingesta, fluids, and tissues from these severely poisoned animals. Urine and ingesta from the upper portion of the gastrointestinal tract, containing from 1 to 20 ppm of cantharidin, were the most satisfactory samples for diagnosing toxicosis. Beetle preparations also were assayed and found to contain widely varying amounts of cantharidin (0.89% to 5.40% of dry weight). Blood chemical analyses on sera and urine samples from the sheep and pony indicated renal dysfunction.

1,3 Butanediol treatment of ethylene glycol toxicosis in dogs.

In order to assess the therapeutic value of 1,3 butanediol in ethylene glycol toxicosis, mixed-bred dogs were given an oral dose of commercial antifreeze at 6 ml/kg of body weight (0 hour) and treated (IV) 7 times at 6-hour intervals with 5.5 ml/kg of body weight 1,3 butanediol solution (20% in physiological saline solution) beginning at 8, 12, and 21 hours. Serum glycolic acid concentration was quantitated by high-pressure liquid chromatography. Three dogs that were given ethylene glycol, but no 1,3 butanediol treatment, died with elevated serum glycolic acid concentrations. Five dogs were given ethylene glycol and 1,3 butanediol treatment. Of 2 dogs treated at 8 hours, 1 survived and 1 died at 39 hours; 1 treated at 12 hours and 1 treated at 21 hours survived; 1 dog died soon (27 hours) after treatment was initiated at 21 hours. Four of the 5 dogs had dramatically decreased serum glycolic acid concentrations after 1,3 butanediol treatment, indicating its effectiveness in inhibiting alcohol dehydrogenase-dependent glycolic acid formation in vivo.

Analytical evaluation of urinary excretion of furosemide in barrows.

Market weight barrows were dosed with furosemide IM or orally, and urine (5 barrows) and blood (1 barrow) samples were collected. Extraction procedures and high-performance liquid chromatographic conditions were modified and adapted for analysis of porcine samples. Furosemide was cleared rapidly from plasma, but could be detected in urine 4 to 5 days after administration, using fluorescent or ultraviolet detection. Fluorescent detection was the preferred method for screening for furosemide. Approximately 12% of furosemide was excreted as the glucuronide conjugate.

Etiologic agents, incidence, and improved diagnostic methods of cantharidin toxicosis in horses.

In addition to the 3-striped blister beetles (Epicauta temexa and E occidentalis), other sources of equine cantharidin toxicosis were identified at the Texas Veterinary Medical Diagnostic Laboratory and included E albida and E attrivittata and the previously incriminated E pardalis and E pennsylvanica. Improved methods for diagnosing cantharidin or blister beetle toxicosis involve partial purification of urine and gastric content extracts, using silica cartridges, followed by analysis, using capillary gas chromatography/mass spectrometry. During a 26-month period, 53 episodes of cantharidin toxicosis in horses were confirmed at our diagnostic laboratory. Concentrations of cantharidin in urine and gastric contents ranged from 0.0003 to 3.50 micrograms/g. Peak incidences were observed in late summer and early fall.

Determination of brodifacoum and bromadiolone residues in rodent and canine liver.

A method to determine residue concentrations of anti-coagulant rodenticides, brodifacoum (BF) and bromadiolone (BD) in liver was developed, using gas chromatography/mass spectrometry. Nine dogs were given 1.1 mg of BF/kg of body weight, PO, in polyethylene glycol 400, one time. Rats were fed BF or BD (via commercial baits) in amounts from 0.28 to 11.25 mg/kg over 1- to 4-day periods. Fresh liver samples were collected at necropsy from all rats and 3 dogs, ground with Na2SO4, and extracted with CHCl3:MeOH (9:1). After evaporation and silica cartridge purification were performed, residues were oxidized with a 0.16M chromic acid solution, and an oxidation product (4-bromobenzoic acid) was partitioned into CHCl3. The methylated derivative (port derivatization with trimethylanilinium hydroxide) was assayed, using gas chromatography/mass spectrometry. Bromadiolone was detected in livers from rats given greater than 6 mg of BD/kg of body weight, but not in livers of rats given 1.25 mg of BD/kg. In contrast, BF was detected (with one exception) in livers from dogs (given 1.1 mg of BF/kg) and from rats given high (11.25 mg of BF/kg) and low (0.28 mg of BF/kg) doses. This protocol, which does not differentiate between BF and BD because of the formation of a common product after chromic acid oxidation, was used to diagnose anticoagulant toxicosis in 3 dogs, 1 human being and 1 llama naturally poisoned.

Experimentally induced Cassia roemeriana poisoning in cattle and goats.

The dried, ground aerial portions of the plant Cassia roemeriana were administered to each of seven calves at a dosage of 10 g/kg of body weight/day for 2 to 10.5 days and to each of six goats at a dosage of 10 g/kg/day for 5 days or 5 or 7 g/kg/day for 23 to 25 days. Experimentally induced C roemeriana poisoning in both species resulted in hepatopathic poisoning characterized by a brief survival period (3.9 to 7.9 days), moderate-to-severe hepatocellular damage, and little or no evidence of injury to skeletal muscle or resulted in myopathic poisoning characterized by a longer period of survival, mild-to-severe skeletal myopathy, and mild hepatocellular injury. The minimal dosage that induced hepatopathic poisoning (also the minimal lethal dosage) was 10 g/kg/day for 3 days in calves and for 5 days in goats. The minimal dosage that induced the myopathic syndrome (as determined by the earliest increase in serum creatine kinase activity) was 10 g/kg/day for 6 days for calves and 5 g/kg/day for 10 to 16 days for goats.

Evaluation of an analytical method for the diagnosis of cantharidin toxicosis due to ingestion of blister beetles (Epicauta lemniscata) by horses and sheep.

An analytical method for cantharidin, using high performance liquid chromatography, was applied to field specimens from horses and sheep with blister beetle (Epicauta lemniscata) poisoning. Stomach content and urine proved to be valuable aids in diagnosis. One incident of cantharidin toxicosis in ruminants (sheep) was confirmed.

Clinical signs and chemical confirmation of 4-aminopyridine poisoning in horses.

4-Aminopyridine poisoning in horses was diagnosed. Specific methods, utilizing thin-layer and high-performance liquid chromatography, were developed for determining the compound in stomach contents and corn bait. The lethal dose was estimated at 2 to 3 mg/kg of body weight.

Photosensitization of cattle in southeast Texas: identification of phototoxic activity associated with Cooperia pedunculata.

A microbiological assay (Candida albicans) was used to screen plants in southeast Texas where bovine photosensitization (PS) of unknown cause was a recurring problem. Phototoxic activity was identified associated with dead leaf tips of Cooperia pedunculata, a native, perennial forb of the Amaryllis family (Amaryllidaceae) from central, southeast, and south Texas and parts of Mexico. A syndrome compatible with naturally occurring PS in cattle was induced in laboratory mice after oral administration of dead leaf material from C pedunculata. Availability and phototoxic activity of dead leaf material of C pedunculata corresponded with occurrence of PS in cattle. Seemingly, C pedunculata was involved in recurring PS.

Waterfowl mortality caused by aflatoxicosis in Texas.

Waterfowl mortality caused by aflatoxicosis occurred in two separate areas in Texas during the 1977-78 wintering season. The first outbreak occurred in snow geese (Anser caerulescens) on the Gulf Coast prairies, followed by an outbreak in mallards (Anas platyrhynchos) in the north-central portion of the state. Aflatoxin B1 levels in geese were 500 ng/g (dry weight). Aflatoxin B1 levels in the second mortality were 10-250 ng/g (dry weight). The exact source of the toxin was not demonstrated in the first outbreak, but in the second outbreak was traced to waste peanuts, which constituted a major portion of the diet of wintering waterfowl in north-central Texas. Aflatoxin B1 levels in the field peanuts collected in the general areas were 110 ng/g.

Benzene hexachloride poisoning in cattle.

Of 174 cattle dipped in an emulsified preparation of benzene hexachloride labeled for plant use, 18 were fatally poisoned. The preparation contained 0.14% active ingredient, gamma isomer of benzene hexachloride (BHC), a normally safe concentration for cattle. Analyses revealed 0.08% gamma BHC in the used dip and 0.5, 7.9, and 198 ppm in liver, kidney, and hairskin specimens, respectively, from a fatally affected cow. Microscopic examination of the freshly prepared dip demonstrated emulsion droplets ranging from 5 to 60 mu in diameter.

Bovine abortion and death associated with consumption of aflatoxin-contaminated peanuts.

Approximately 12% of a herd of 68 crossbred cows aborted third-trimester fetuses after consuming moldy peanuts for 4 days. Further investigation revealed that less than 20% of the herd had access to this supplemental feed. Results of serum biochemical analysis indicated liver damage in the affected cows. All of these cows died within 8 days of aborting. The peanuts contained 77 micrograms aflatoxin B1/g, as determined by liquid chromatography. Tissues were submitted from 1 cow, and liver contained 5 ng aflatoxin B1/g. Results of other laboratory tests were negative for common toxins and abortifacients.

Catastrophic death losses in a dairy herd attributed to type D botulism.

Clostridium botulinum type D intoxication was diagnosed as the cause of death of 42 of 67 lactating cows in a southeast Texas dairy herd over an 11-day period. By necessity, the diagnosis was based on clinicopathologic findings, as the toxin could not, by standard laboratory tests, be demonstrated in affected cattle. The predominant clinical findings were hindlimb weakness/ataxia rapidly progressing to persistent recumbency. Affected cattle were alert until just before death, which occurred without notable agonal movements or respirations after 6 to 72 hours' recumbency. Abnormal laboratory findings included neutrophilic leukocytosis (all affected cattle), proteinuria (most affected cattle), slight elevations of serum aspartate transaminase and low serum inorganic phosphorus (some affected cattle), and patchy areas of hyperemia/congestion of the mucosa in the small intestine (postmortem examination of 3 affected cattle). This report confirms the findings of others with regard to the difficulty of demonstrating the causative toxin in C botulinum type D-intoxicated cattle and presents available information on the clinicopathologic features of this intoxication that may aid in the differentiation of this condition from other causes of down cows.

Toxic plants. What the horse practitioner needs to know.

Horses poisoned by a plant may show a myriad of clinical signs. Awareness of the poisonous plants growing in a given area and those that are likely to appear in hay and their associated clinical signs can be instrumental in making diagnoses. More importantly, the information can be shared with clients to help prevent plant poisonings in horses.