Vasoactive Effects of Acute Ergot Exposure in Sheep.

Ergotism is a common and increasing problem in Saskatchewan's livestock. Chronic exposure to low concentrations of ergot alkaloids is known to cause severe arterial vasoconstriction and gangrene through the activation of adrenergic and serotonergic receptors on vascular smooth muscles. The acute vascular effects of a single oral dose with high-level exposure to ergot alkaloids remain unknown and are examined in this study. This study had two main objectives; the first was to evaluate the role of α1-adrenergic receptors in mediating the acute vasocontractile response after single-dose exposure in sheep. The second was to examine whether terazosin (TE) could abolish the vascular contractile effects of ergot alkaloids. Twelve adult female sheep were randomly placed into control and exposure groups (n = 6/group). Ergot sclerotia were collected and finely ground. The concentrations of six ergot alkaloids (ergocornine, ergocristine, ergocryptine, ergometrine, ergosine, and ergotamine) were determined using HPLC/MS at Prairie Diagnostic Services Inc., (Saskatoon, SK, Canada). Each ewe within the treatment group received a single oral treatment of ground ergot sclerotia at a dose of 600 µg/kg BW (total ergot) while each ewe in the control group received water. Animals were euthanized 12 h after the treatment, and the pedal artery (dorsal metatarsal III artery) from the left hind limb from each animal was carefully dissected and mounted in an isolated tissue bath. The vascular contractile response to phenylephrine (PE) (α1-adrenergic agonist) was compared between the two groups before and after TE (α1-adrenergic antagonist) treatment. Acute exposure to ergot alkaloids resulted in a 38% increase in vascular sensitivity to PE compared to control (Ctl EC50 = 1.74 × 10-6 M; Exp EC50 = 1.079 × 10-6 M, p = 0.046). TE treatment resulted in a significant dose-dependent increase in EC50 in both exposure and control groups (p < 0.05 for all treatments). Surprisingly, TE effect was significantly more pronounced in the ergot exposed group compared to the control group at two of the three concentrations of TE (TE 30 nM, p = 0.36; TE 100 nM, p < 0.001; TE 300 nM, p < 0.001). Similar to chronic exposure, acute exposure to ergot alkaloids results in increased vascular sensitivity to PE. TE is a more potent dose-dependent antagonist for the PE contractile response in sheep exposed to ergot compared to the control group. This study may indicate that the dry gangrene seen in sheep, and likely other species, might be related to the activation of α1-adrenergic receptor. This effect may be reversed using TE, especially at early stages of the disease before cell death occurs. This study may also indicate that acute-single dose exposure scenario may be useful in the study of vascular effects of ergot alkaloids.

Use of Integrative Interactomics for Improvement of Farm Animal Health and Welfare: An Example with Fescue Toxicosis.

Rapid scientific advances are increasing our understanding of the way complex biological interactions integrate to maintain homeostatic balance and how seemingly small, localized perturbations can lead to systemic effects. The 'omics movement, alongside increased throughput resulting from statistical and computational advances, has transformed our understanding of disease mechanisms and the multi-dimensional interaction between environmental stressors and host physiology through data integration into multi-dimensional analyses, i.e., integrative interactomics. This review focuses on the use of high-throughput technologies in farm animal research, including health- and toxicology-related papers. Although limited, we highlight recent animal agriculture-centered reports from the integrative multi-'omics movement. We provide an example with fescue toxicosis, an economically costly disease affecting grazing livestock, and describe how integrative interactomics can be applied to a disease with a complex pathophysiology in the pursuit of novel treatment and management approaches. We outline how 'omics techniques have been used thus far to understand fescue toxicosis pathophysiology, lay out a framework for the fescue toxicosis integrome, identify some challenges we foresee, and offer possible means for addressing these challenges. Finally, we briefly discuss how the example with fescue toxicosis could be used for other agriculturally important animal health and welfare problems.

Identification of bovine prolactin in seminal fluid, and expression and localization of the prolactin receptor and prolactin-inducible protein in the testis and epididymis of bulls exposed to ergot alkaloids.

The objectives of this study were to determine (1) the presence and expression levels of bovine prolactin receptor (PRLR) and prolactin-inducible protein (PIP) in bovine testis and epididymis, and (2) the presence and concentrations of prolactin (PRL) present in seminiferous fluid in bulls consuming diets with (E+) or without (E-) ergot alkaloids. Bulls (n = 8) were sacrificed after 126 days (group A) of E+ or E- treatment or 60 days after all bulls (n = 6) were switched to the E- ration (group B). End point and real-time quantitative reverse transcription-polymerase chain reaction and immunohistochemistry were conducted on testis and epididymis samples to establish the presence and relative expression of PRLR and PIP. Seminal fluid samples obtained from bulls consuming E- and E+ diets were subjected to RIA for PRL. Both PIP and PRLR were present in testis and epididymis as determined by reverse transcription-polymerase chain reaction and immunohistochemistry. Prolactin-inducible protein mRNA abundance was affected by time of slaughter in testis and epididymis head, respectively (P < 0.05). Prolactin receptor mRNA expression was affected by time of slaughter in the epididymis (P < 0.05) and differed in testis samples because of treatment (P < 0.05). Radioimmunoassay establishes the presence of PRL in seminal fluid; however, differences in the concentration of PRL over two separate studies were inconsistent, possibly because of differences in diet. The presence and localization of the PRLR are consistent with expression data reported for other species, and the presence of PIP and PRL in seminal fluid is consistent with data generated in humans.

Board-invited review: St. Anthony's Fire in livestock: causes, mechanisms, and potential solutions.

After a brief history of ergot alkaloids and ergotism, this review focuses on the metabolism and mechanisms of action of the ergot alkaloids. The authors provide models of how these alkaloids afflict grazing livestock under complex animal-plant/endophyte-environmental interactions. Alkaloid chemistry is presented to orient the reader to the structure-function relationships that are known to exist. Where appropriate, the medical literature is used to aid interpretation of livestock research and to provide insight into potential modes of action and alkaloid metabolism where these are not known for livestock. In closing the paper, we discuss management of ergot alkaloid intoxication in livestock and future research needs for this field of study.

Effects of short-term heat stress on endophytic ergot alkaloid-induced alterations in rat hepatic gene expression.

Exposure to ergot alkaloids in endophyte-infected fescue (E+) is associated with impaired animal productivity, especially during heat stress, which is commonly referred to as fescue toxicosis. To elucidate the pathogenesis of this condition, the effects of short-term heat stress (HS) on hepatic gene expression in rats exposed to endophytic ergot alkaloids were evaluated. Rats implanted with telemetric transmitters to continuously measure core temperature were fed an E+ diet and maintained under thermoneutral (TN) conditions (21 degrees C) for 5 d, followed by TN or 31 degrees C (HS) conditions for 3 d. Feed intake (FI) and BW were monitored daily. The E+ and HS-induced alterations in hepatic genes were evaluated using DNA microarrays and PCR analyses. Hepatic antioxidant enzyme activities, as well as the incidence of apoptosis, were determined. As expected, intake of E+ reduced FI and BW from pretreatment levels under TN conditions, with greater reductions during short-term HS. Genes involved in gluconeogenesis and apoptosis were upregulated, whereas genes associated with oxidative phosphorylation, xenobiotic metabolism, antioxidative mechanisms, immune function, cellular proliferation, and chaperone activity were all downregulated with short-term HS. Hepatocytic apoptosis was increased and antioxidant enzyme activity decreased in the livers of rats exposed to HS. The hypothesized, exacerbating effects of HS on the direct, endophytic toxin-related and indirect, reduced caloric intake-associated alterations in hepatic gene expression were clearly demonstrated in rats and may help to elucidate the pathogenesis of fescue toxicosis in various animal species.

Endocrine alterations associated with ergopeptine alkaloid exposure during equine pregnancy.

Ergopeptine alkaloid exposure is common in pregnant mares. Many mares live in geographic areas where Neotyphodium coenophialum-infected tall fescue is the dominant grass in pastures and hay. A variety of grasses and cereal grains can be infected by Claviceps purpurea, and fungal sclerotia can contaminate forage and especially ground and pelleted feed. An understanding of the endocrine alterations associated with ergopeptine alkaloid exposure during pregnancy is necessary for the diagnosis of potential exposure to these compounds and for eective prophylaxis and therapy.

Milk production in Angus, Brahman, and reciprocal-cross cows grazing common bermuda grass or endophyte-infected tall fescue.

Milk yield and quality were measured on 139 Angus, Brahman, and reciprocal-cross cows grazing common bermuda grass or endophyte-infected tall fescue for 4 yr to evaluate interactions of direct and maternal breed effects and heterosis for these traits with forage environment. Milk yield was estimated by method of milking machine, and milk fat, protein, and somatic cell counts were evaluated in a commercial dairy laboratory. Monthly estimates were made beginning on an average d 61 of lactation and continued monthly for six estimates in 3 yr and five estimates in 1 yr. Data were averaged over month within year, and the model included sire breed, sire in sire breed, dam breed, forage, and age averages. Somatic cell counts were transformed using natural logarithms prior to analyses. Forage effects for milk yield were dissimilar among sire breed x dam breed subclasses (P < .10), resulting in higher levels of heterosis on common bermuda grass than on tall fescue. Maternal breed effects for milk yield favored Angus on bermuda grass (P < .05) but not on tall fescue, whereas direct breed effects were similar on both forages and favored Brahman. Milk fat was reduced on tall fescue compared to bermuda grass by an average of .6% (P < .01), and direct breed effects were similar across forages and averaged 1.04% (P < .01) in favor of Brahman. Heterosis and maternal breed effects for milk fat were not important. There was little evidence of direct and maternal breed effects or heterosis for milk protein or somatic cell counts. These data suggest that heterosis for milk yield is larger on common bermuda grass than on tall fescue and that grazing endophyte-infected tall fescue is detrimental to milk fat.

The effects of fescue toxicosis on beef cattle productivity.

Consumption of tall fescue forage infested with the endophytic fungus Acremonium coenophialum can result in a condition termed "fescue toxicosis," which is characterized by decreased weight gains, milk production, conception, and serum prolactin and an inability to dissipate body heat by beef cattle. These decreases in productivity have been estimated to cost beef producers more than $600 million annually. The unthrifty appearance of cattle consuming endophyte-infected tall fescue (E+) is most evident during periods of environmental heat or cold stress, suggesting an interaction with environmental conditions. Without temperature stress, cows consumed similar amounts of E+ and endophyte-free tall fescue (E-). But, when temperatures exceeded 32 degrees C, cows that grazed E+ consumed less forage than cows that grazed E-. After removal of E+ from the diet of steers, compensatory growth was observed, indicating no long-term negative effects of E+. A decrease in serum prolactin but apparently little change in other hormones has been measured after consumption of E+. Under periods of heat stress, animals had reduced ability to dissipate body heat, and blood flow to peripheral (rib skin), core-body (duodenum colon), and brain (cerebellum) tissues was decreased. Due to the depressive effects of E+ on prolactin and heat dissipation, dopamine antagonist therapy has been used in an attempt to rectify these changes. Dopamine antagonists have increased serum prolactin but there is limited evidence to suggest an improvement in heat dissipation. Experiments to elucidate the effects of E+ on alpha-1 and -2 adrenergic receptors may also offer insights into developing strategies to overcome the negative effects of E+ consumption. Preliminary results suggest that injections of an alpha-1 adrenergic antagonist (prazosin) in rats fed E+ under 32 degrees C conditions increased DM intake and reduced rectal temperature.

[The chemical-toxicological analysis of ergot alkaloids]. / Khimiko-toksikologicheskii analiz alkaloidov sporyn'i.

Russian and foreign literature dealing with this topic is reviewed. Brief descriptions of methods of examinations starting from color tests and up to high-pressure liquid chromatography and mass spectrometry are presented in a chronological order. Clinical and toxicological characteristics of some ergot alkaloids are described.

Further studies concerning the toxicity of ingested ergot sclerotia (Claviceps purpurea) to young and growing pigs.

In three experiments pigs were offered diets contaminated with milled ergot sclerotia. In a metabolism study 0-5 per cent (w/w) of American wheat ergot decreased urinary nitrogen losses and improved the efficiency of nitrogen retention. When early weaned piglets ingested a diet contaminated with 2-5 per cent of the ergot, feed intake was reduced and growth rate decreased. Reduction in growth rate was also evident in growing pigs given a diet containing ergotised English wheat. These latter pigs had lesions in stomach, intestine and liver, consistent with a toxic insult. With respect to feed intake and growth these results agreed with those of an earlier study, although a toxic response was produced by a lower dosage rate and there was additional internal evidence of toxicity. Improved nitrogen retention was contrary to earlier evidence at higher dose rates but may merely reflect a tonic action which, at higher doses, becomes toxic.