Ergot alkaloids reduce circulating serotonin in the bovine

Ergot alkaloids can interact with several serotonin (5-hydroxytryptamine [5-HT]) receptors provoking many physiological responses. However, it is unknown whether ergot alkaloid consumption influences 5-HT or its metabolites. Thus, two experiments were performed to evaluate the effect of ergot alkaloid feeding on 5-HT metabolism. In exp. 1, 12 Holstein steers (260 ± 3 kg body weight [BW]) were used in a completely randomized design. The treatments were the dietary concentration of ergovaline: 0, 0.862, and 1.282 mg/kg of diet. The steers were fed ad libitum, kept in light and temperature cycles mimicking the summer, and had blood sampled before and 15 d after receiving the treatments. The consumption of ergot alkaloids provoked a linear decrease (P = 0.004) in serum 5-HT. However, serum 5-hydroxytryptophan and 5-hydroxyindoleacetic acid did not change (P > 0.05) between treatments. In exp. 2, four ruminally cannulated Holstein steers (318 ± 3 kg BW) were used in a 4 × 4 Latin square design to examine the difference between seed sources on 5-HT metabolism. Treatments were: control—tall fescue seeds free of ergovaline, KY 32 seeds (L42-16-2K32); 5Way—endophyte-infected seeds, 5 way (L152-11-1739); KY31—endophyte-infected seeds, KY 31 (M164-16-SOS); and Millennium—endophyte-infected seeds, 3rd Millennium (L108-11-76). The endophyte-infected seed treatments were all adjusted to provide an ergovaline dosage of 15 μg/kg BW. The basal diet provided 1.5-fold the net energy requirement for maintenance. The seed treatments were dosed directly into the rumen before feeding. The experiment lasted 84 d and was divided into four periods. In each period, the steers received seeds for 7 d followed by a 14-d washout. Blood samples were collected on day 0 (baseline) and day 7 for evaluating the treatment response in each period. A 24 h urine collection was performed on day 7. Similar to exp. 1, serum 5-HT decreased (P = 0.008) with the consumption of all endophyte-infected seed treatments. However, there was no difference (P > 0.05) between the infected seeds. The urinary excretion of 5-hydroxyindoleacetic acid in the urine was not affected (P > 0.05) by the presence of ergot alkaloids. In conclusion, the consumption of ergot alkaloids decreases serum 5-HT with no difference between the source of endophyte-infected seeds in the bovine.     

Histopathology and a real-time PCR assay for detection of Bonamia ostreae in Ostrea edulis cultured in western Canada

Bonamia ostreae is an intracellular haplosporidian parasite in European flat oysters Ostrea edulis that occurs on both coasts of the United States and causes significant mortality in Europe. Canada was considered free of B. ostreae until 2004, when it was first detected in O. edulis obtained for laboratory study from a western Canadian oyster farm. Bonamia ostreae was confirmed in O. edulis at the index farm in November 2004 using histopathology, conventional polymerase chain reaction (PCR) assays, restriction fragment length polymorphism (RFLP) analysis, and sequencing of the PCR product. Archived samples of European flat oysters obtained from the index farm between 1999 and 2004 (n = 343) were re-examined and all samples collected before 2003 (n = 306) were confirmed negative for B. ostreae by histopathology (n = 306) and PCR (n = 62). In archived samples from 2003, B. ostreae was detected in 3 of 37 O. edulis by histopathology (n = 2) and/or PCR (n = 2). Also, records indicate that B. ostreae was not detected in O. edulis (n = 348) from five other locations in western Canada between 1986 and 2000. To better understand the distribution and prevalence of B. ostreae in western Canada, 607 oysters from the index farm and 2 additional farms were sampled in the summer of 2005. All 3 farms had been stocked with O. edulis spat from the State of Washington, USA, where B. ostreae is endemic. Samples were analyzed by histopathology and a new real-time PCR that amplifies a 68-bp target DNA fragment. B. ostreae was detected in all three locations, with prevalence ranging from 0.5 to 11.1%. Diagnostic sensitivity of the real-time PCR method was consistently greater than histopathology. Also, preliminary evidence supports the conclusion that real-time PCR on paraffin sections is more sensitive than histopathology; B. ostreae DNA was confirmed in 4 oysters by real-time PCR on paraffin-embedded tissues (and independently confirmed on unfixed tissues) that was not detected by histopathology. As a result of these findings, O. edulis spat are no longer allowed to be imported from endemic areas into Canada.