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The Oregon State University Colleges of Veterinary Medicine and Agricultural Sciences instituted the Endophyte Service Laboratory to aid in diagnosing toxicity problems associated with cool-season grasses in livestock. The endophyte (Neotyphodium coenophalum) present in tall fescue (Festuca arundinacea) produces ergopeptine alkaloids, of which ergovaline is the molecule used to determine exposure and toxicity thresholds for the vasoconstrictive conditions "fescue foot" and "summer slump". Another vasoconstrictive syndrome, "ergotism," is caused by a parasitic fungus, Claviceps purpurea, and its primary toxin, ergotamine. "Ryegrass staggers" is a neurological condition that affects livestock consuming endophyte (Neotyphodium lolii)- infected perennial ryegrass (Lolium perenne) with high levels of lolitrem B. HPLC-fluorescent analytical methods for these mycotoxins are described and were used to determine threshold levels of toxicity for ergovaline and lolitrem B in cattle, sheep, horses, and camels. In addition, six clinical cases in cattle are presented to illustrate diagnosis of these three diseases.

Mineral implications in health issues of captive rhinos have received much attention lately. This study was undertaken to establish reference values for the mineral status of rhinos. Serum/plasma samples of free-ranging black (Diceros bicornis) and white (Ceratotherium simum) rhinos and of captive black, white, Indian (Rhinoceros unicornis), and Sumatran (Dicerorhinus sumatrensis) rhinos, as well as liver tissue samples of captive black, white, and Indian rhinos were analyzed for mineral content. Circulating mineral levels of free-ranging animals were subject to variation according to region. In free-ranging animals, high molybdenum (Mo) values compared to horse normals were striking. Captive animals displayed even higher circulating Mo concentrations. The significance of iron (Fe) overload in captive specimens of the browsing rhinos (black and Sumatran) was confirmed. Hepatic Fe levels increased in blacks with age. Although this Fe overload is suspected to be linked with diets, the data indicate that this is not due solely to an excessive dietary Fe supply. Whereas the grazing species (white and Indian) had high liver copper (Cu) levels, the browsing species had low to marginal liver Cu concentrations. Liver concentrations of K, Mg, Co, and Mo increased with age in captive black rhinos. Additional findings include high circulating Se levels in all rhino species. Future research should be directed at investigating factors leading to high Fe levels, and at investigating Cu metabolism in captive rhinoceros species.

Ergot-induced disease in humans was known long before Biblical times and has been the root cause for countless human epidemics spanning from the early fourteenth century to the late sixteenth century. In contrast, many of these same ergot alkaloids have been utilized for their medicinal properties to mitigate migraine headaches and have had indications as anti-carcinogens. Although ergot alkaloids have been used for centuries by humans, basic pharmacokinetic data has not been documented for clinical disease in livestock. Consequently, a threshold dose and accurate dose-response data have yet to be established. Throughout the past several years, new detection techniques have emerged to detect these alkaloids at the parts per billion (ppb) level which has allowed for new efforts to be made with respect to determining threshold levels and making accurate clinical diagnoses in affected animals. This perspectives article provides a critical initial step for establishing a uniform interpretation of ergot toxicosis from limited existing data.

Intravenous dimethyl sulfoxide (DMSO) has been utilized as a treatment for a number of acute inflammatory conditions and for cerebral edema following trauma in experimental and clinical cases. 2,5,9, 11,12,15,16,18,20.2t.23,2s In humans, DMSO at 1 g/kg, has been found to be more effective than mannitol, urea, cortisone or barbiturates in treating severe head trauma. TM This treatment is often the most economical and effective way of treating brain edema in acute equine neurological diseases, z In the horse, recommended intravenous dosages range from 0.1 g/kg to 4 g/kg in varying concentrations of up to 40% DMSO. l'2'r'1s'21 While the most commonly used dose is 1 g/kg 1,2,19,~ the adverse effects of DMSO given too rapidly at any dose have not been well documented in horses. Adverse effects relative to DMSO have been reported in human beings and a number of animal species. 4,8 These include intravascular hemolysis, hemoglobinuria, hyperosmolality, and transient increase in respiratory rate. Clinical signs associated with near lethal doses in experimental animals were spontaneous motor activity, tremors, myasthenia

Ergot alkaloids are mycotoxins generated by grass and grain pathogens such as Claviceps, for example. Ergot alkaloid-poisoning syndromes, such as tall fescue toxicosis from endophyte-infected tall fescue grass, are important veterinary problems for cattle, horses, sheep, pigs and chickens, with consequent impact on food, meat and dairy industries. Damage to livestock is of the order of a billion dollars a year in the United States alone. HPLC with UV and fluorescence detection are the predominant means of ergot alkaloid determination, with focus on quantitation of the marker compound ergovaline, although ELISA methods are undergoing investigation. These techniques are excellent for rapid detection, but of poor specificity in defining new or poorly characterized ergot alkaloids and related compounds. This paper demonstrates the facility of using electrospray(+) mass spectrometry with multiple reaction monitoring (MRM) detection during chromatographic examination of ergot alkaloid standards of lysergic acid, lysergol, ergonovine, ergovaline, ergotamine, ergocornine, ergocryptine and ergocrystine by HPLC. Ergoline-8 position epimers could be separated on the gradient HPLC system for ergocornine, ergocrystine and ergonovine and appeared as shoulders for ergotamine and ergovaline; epimers generally showed different patterns of relative intensity for specific MRM transitions. There was reasonable correspondence between retention of standards on the 2-mm ESI(+)MS phenyl-hexyl-based reverse phase column and those on the 4-mm C18-based column. Since up to 10% of clinical cases involving toxin exposure display unidentified chromatographic peaks, 11 samples of feed components associated with such cases were studied with developed MRM methods to attempt elucidation of crucial components if possible. Ergotamine appeared in all, ergovaline appeared in five and ergocornine appeared in six; ergonovine, ergocryptine, ergocrystine and lysergol also appeared in several. In addition, molecular weights of compounds newly revealed by mass spectrometry suggested ergosine, ergostine and ergoptine in four samples, for which standards were not available. Dehydrated products of ergotamine, ergocrystine and ergocornine were discovered, along with dihydrogenated ergocrystine and ergocryptine in seven of the samples, and the issue was raised as to whether dehydration was strictly an instrument-derived artifact. Finally, five of the samples, along with fescue seed standard, evidenced one or more of 14 new ergot alkaloids ranging in size from 381 to 611 molecular weight and with key mass spectral characteristics of ergot alkaloids, specifically the pair of peaks m/z 223 and 208, corresponding to the ergoline ring system and its demethylated variant, respectively. It is anticipated that findings such as these will provide impetus to future development of analytical methodology for these heretofore relatively rare ergot alkaloid species.

Objective—To determine whether iontophoretic administration of dexamethasone to horses results in detectable concentrations in synovial fluid, plasma, and urine. Animals—6 adult mares. Procedure—Iontophoresis was used to administer dexamethasone. Treatments (4 mA for 20 minutes) were administered to a tarsocrural joint of each mare. The drug electrode contained 3 ml of dexamethasone sodium phosphate at a concentration of 4 or 10 mg/ml. Samples of synovial fluid, blood, and urine were obtained before and 0.5, 4, 8, and 24 hours after each treatment. All samples were tested for dexamethasone using an ELISA. Synovial fluid also was evaluated for dexamethasone, using high-performance liquid chromatography. Results—The lower and upper limits of detection for dexamethasone in synovial fluid with the ELISA were 0.21 and 1.5 ng/ml, respectively. Dexamethasone administered at a concentration of 10 mg/ml was detected by the ELISA in synovial fluid of 5 mares from 0.5 to 24 hours and in urine of 4 mares from 0.5 to 8 hours after each treatment, but it was not detected in plasma. Mean synovial fluid concentration of dexamethasone was 1.01 ng/ml. Dexamethasone administered at a concentration of 4 mg/ml was detected by the ELISA in urine of 2 mares at 0.5 and 4 hours after treatment, but it was not detected in synovial fluid or plasma. Conclusion and Clinical Relevance—Iontophoresis cannot be considered an effective method for delivery of dexamethasone to synovial fluid of horses, because drug concentrations achieved in this study were less than therapeutic concentrations. (Am J Vet Res 2002;63:11–14)

Objective—To determine the serum concentrations and sedative effects of fentanyl after transdermal administration at 3 dosages in llamas. Animals—9 healthy adult female llamas (mean age, 8 ± 3 years; mean weight, 150 ± 18 kg). Procedure—Llamas were allocated to 1 of 3 groups (3 llamas/group). Fentanyl patches (each providing transdermal delivery of 75 µg of fentanyl/h) were placed on shaved areas of the antebrachium of all llamas. In group 1, llamas were treated with 1 patch (anticipated fentanyl dosage, 75 µg/h). In group 2, llamas were treated with 2 patches (anticipated fentanyl dosage, 150 µg/h). In group 3, llamas were treated with 4 patches (anticipated fentanyl dosage, 300 µg/h). For each llama, the degree of sedation was assessed by use of a subjective scoring system and a blood sample was collected for determination of serum fentanyl concentration at 12, 24, 36, 48, 60, and 72 hours after patch placement. Results—Following the placement of 4 patches, mean ± SD serum fentanyl concentration in group 3 llamas reached 0.3 ± 0.08 ng/mL within 12 hours. This concentration was sustained for 72 hours. In group 2, application of 2 patches provided inconsistent results; in group 1, application of 1 patch rarely provided measurable serum fentanyl concentrations. No llamas became sedated at any time. Conclusions and Clinical Relevance—Results suggest that application of four 75 µg/h fentanyl patches provides consistent, sustained serum fentanyl concentrations without sedation in llamas. However, the serum concentration of fentanyl that provides analgesia in llamas is not known. (Am J Vet Res 2005;66:907–909)

The essential oil extracts of western juniper oil (Juniperus occidentalis) and Port-Orford-cedar oil (Chamaecyparis lawsoniana) were evaluated for possible dermal toxic effects on mice and rabbits. Mice were tested for their response to both extracts utilizing a local lymph node assay. Western juniper oil extract at 0.5% and 5% concentrations did not show a stimulation index (SI) greater than normal (3.0); however, a 50% concentration did show a positive response at 3.3. Port-Orford-cedar oil extract did not show a positive response at concentrations of 0.5%, 5% or 50%. An acute dermal irritation study using rabbits had a primary irritation index (PII) of 3.3 with 100% Port-Orford-cedar oil extract. This was reduced to a PII of 0.625 when diluted 1:1 with olive oil. Undiluted western juniper oil extract had a PII score of 2.7. While a 5.0% solution had a PII score of 0.3, a 0.5% solution of western juniper oil was a non-irritant. It would appear that animals bedded on wood shavings have contact with essential oils at concentrations far less than the 2% maximum by weight obtained by steam distillation extraction. These concentrations did not elicit a hypersensitivity response.

PIERCY, R. J., K. W. HINCHCLIFF, P. S. MORLEY, R. A. DISILVESTRO, G. A. REINHART, S. L. NELSON, JR., K. E. SCHMIDT, and A. M. CRAIG. Association between vitamin E and enhanced athletic performance in sled dogs. Med. Sci. Sports Exerc., Vol. 33, No. 5, 2001, pp. 826-833. Purpose: To determine the association between prerace plasma vitamin E concentration and performance in sled dogs competing in the 1998 Iditarod Race. Methods: Prerace blood samples were collected from 670 dogs. Samples were analyzed for plasma vitamin E concentration while controlling for selected hematological and biochemical variables and signalment. Starting in teams of 16, exercise consisted of running up to 1159 miles pulling a laden sled and musher via checkpoints. The records of dogs that were withdrawn from the race for health reasons, fatigue, or strategic or technical reasons, and those of dogs that finished the race were analyzed. Multiple logistic regression and Cox proportional hazards analysis were used to determine factors associated with endurance. Multiple linear regression analysis was used to determine factors associated with team speed. Results: A total of 323 dogs (48%) were withdrawn from racing at various distances from the start. Median time to finish for 39 teams was 11.5 d d and the winning time was 9.2 d. Dogs with prerace plasma vitamin E concentrations > 40.7 μg.mL -1 were 1.9 times more likely to finish (P = 0.0006) and had 1.8 times less of a risk of being withdrawn for every mile ran (P = 0.03) than were dogs with plasma vitamin E concentrations between 16.3 and 40.7 μg.mL -1 . Neither a team's mean prerace vitamin E concentration, nor the proportion of dogs within a team with high (>40.7 μg.mL -1 ) vitamin E concentration was associated with team speed. Conclusions: Dogs with higher plasma vitamin E concentrations have enhanced endurance compared with dogs with lower plasma vitamin E concentrations, but the plasma vitamin E status of a team is not associated with team speed.

Exertional rhabdomyolysis (ER) is common in sled dogs, animals with high energy expenditures that consume high fat (60% of ingested calories) diets. Associations between pre-race plasma [vitamin E] and total antioxidant status (TAS) and risk of developing ER were examined in dogs competing in the 1998 Iditarod race. Pre-race blood samples were collected from 750 dogs and a second sample was collected from 158 dogs withdrawn from the race at various times. Plasma creatine kinase activity was used to identify withdrawn dogs with ER. There was no association between pre-race plasma [vitamin E] and risk of development of ER. Dogs that developed ER started the race with higher TAS, but when withdrawn, had lower TAS than unaffected dogs and had similar pre-race [vitamin E] but higher [vitamin E] at time of withdrawal. Hence, the risk of ER in sled dogs is not affected by plasma [vitamin E] before the race.

Sodium cyanate, a neurotoxic chemical in rodents, primates and humans, is implicated in neurodegenerative disorders in protein-deficient populations subsisting in parts of Africa on the cyanogenic plant cassava. The molecular and cellular mechanisms of cyanate neurotoxicity are not understood. This study investigates the effect of sodium cyanate on glutathione (GSH) homeostasis in rodent brain and liver in vitro and in vivo. GSH levels in mouse brain were rapidly, time- and dose-dependently decreased following intraperitoneal administration of 100, 200 or 300 mg/kg sodium cyanate. By contrast, GSH disulfide (GSSG) levels were increased and GSH/GSSG ratios were decreased in a dose-dependent manner in rat brain. Sodium cyanate depleted GSH levels in all regions of mouse brain. Brain glutathione reductase activity was dose-dependently inhibited, while glutathione peroxidase activity was not affected by sodium cyanate. The disruption of GSH homeotasis, as evidenced by reduced tissue GSH/GSSG ratios, likely results from cyanate-induced inhibition of glutathione reductase activity. The results of this study suggest that cyanate neurotoxicity, and perhaps cassava-associated neurodegenerative diseases, are mediated in part by disruption of glutathione homeostasis in neural tissue.

Selected serum constituents were analyzed from 50 adult mallards (Anas platyrhynchos) of both sexes during several stages of reproduction: pre-egg laying, egg laying, incubating, molting, and postreproductive. Similar assays were conducted on sera from ducklings aged 5 to 58 days. Values for total protein (TPR), albumin (ALB), glucose (GLU), gamma-glutamyl transferase (GGT), calcium (CA), phosphorus (PHOS) and magnesium (MG) differed by sex. When all data were combined and analyzed for sex-related differences within each reproductive condition separately, all assays except lactate dehydrogenase (LD-L), cholinesterase (CHE), alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine (CRN) and direct bilirubin (BIDI) differed between sexes during one or more reproductive periods. Each assay showed differences among the various reproductive conditions regardless of gender. The pattern of change differed between sexes. All assays except ALB, GLU, CA and MG showed age-related changes. Lipemia in the sample interfered with all chemistries except TPR, LD-L and CA. Results indicate that when using clinical chemistry as a diagnostic tool in the mallard, age and reproductive condition should be determined in order to compare the data to appropriate control values.

Summary Relationships among hemoglobin (Hb) types and production traits were examined in 294 crossbred ewes from North Country Cheviot, Dorset, Finnsheep and Romney rams, and Suffolk and Columbia-type ewes. Hb BB ewe lambs were youngest at first estrus, while Hb AA ewe lambs were oldest. Ewe lamb fertility was similar for Hb AB and Hb BB ewes, and both were superior to Hb AA ewes. When Finnsheep crossbreds were excluded, frown which Hb BB was absent, Hb BB ewes had higher ewe lamb fertility than did Hb AB ewes. There was a minor advantage of the A over the B allele in ewe lamb prolificacy. Hb BB ewes were highest, Hb AB ewes were intermediate and Hb AA ewes were lowest for average fertility, prolificacy per ewe lambing, lambs born per ewe exposed to mating, total number of lambs weaned and total kilograms of lamb weaned. Grease wool production, staple length, fiber diameter grade, incidence of medullated fibers and incidence of cotted fleeces all were similar across Hb types. Likewise, Hb type did not affect milk production or composition. Ewes with Hb AB had the lowest incidence of footrot; Hb AA and Hb BB ewes were similar. Ewes with Hb AA had the lowest fecal parasite egg counts, while ewes with Hb AB or Hb BB were similar. Ewes with Hb AA also had the lowest incidence of mastitis, Hb BB ewes were intermediate and Hb AB ewes were highest. The combination of overdominance (as for footrot resistance), beneficial effects of the A allele on health-related traits (as for mastitis and parasite resistance) and the beneficial effect of the B allele on reproductive traits, if real, could partly explain the genetic polymorphism commonly reported at the Hb locus in sheep.

Synthetic vitamin E, all-rac-alpha-tocopherol, contains eight different stereoisomers, only one of which, RRR-alpha-tocopherol, occurs naturally. The objective of this investigation was to evaluate the relative abilities of these two vitamin E forms to enrich piglet tissues when fed as alpha-tocopheryl acetates to sows during pregnancy and lactation. alpha-Tocopherol delivery to fetuses and to suckling piglets was monitored by feeding 150 mg each of d(3)-RRR-alpha- and d(6)-all-rac-alpha-tocopheryl acetate to three pregnant sows daily from 7 d before to 7 d after giving birth. Labeled and unlabeled vitamin E concentration ratios were measured in sow plasma and milk, and in piglet (n = 9) plasma and tissues at birth, 7 and 21 d. At birth, despite elevated sow plasma deuterated alpha-tocopherol concentrations, no labeled alpha-tocopherol was detected in piglet plasma or tissues. Sow plasma and milk d(3)-alpha- to d(6)-alpha-tocopherol concentrations were 2:1, leading to a 2:1 ratio in suckling piglet plasma and tissues. In piglets at d 7 compared with birth, most tissues contained a 10-fold increase in total alpha-tocopherol; the highest deuterated vitamin E concentrations were in the liver, followed by the lung, heart, kidney, muscle, intestine and brain. In conclusion, pigs discriminate between RRR- and all-rac-alpha-tocopherol with a preference for RRR-alpha-tocopherol; thus, the official bioequivalence ratio of 1.36:1 RRR- to all-rac-alpha-tocopherol is underestimated. After initiation of suckling, piglet plasma and tissues demonstrated a dramatic increase in vitamin E concentrations, emphasizing the limited placental vitamin E transfer and the importance of milk for enhancing the vitamin E status of the newborn.