Baggs ewes adapt to maternal undernutrition and maintain conceptus growth by maintaining fetal plasma concentrations of amino acids

Adequate delivery of AA is essential for normal fetal growth and development. Recently, we reported that when ewes from the University of Wyoming flock (farm flock with adequate nutrition) were fed 50% (nutrient-restricted) or 100% (control-fed) of the NRC-recommended nutrient requirements between d 28 and 78 of gestation, fetal weights as well as concentrations of most AA in maternal and fetal blood were substantially reduced in nutrient-restricted vs. control-fed pregnancies. The current study utilized Baggs ewes, which were selected under a markedly different production system (range flock with limited nutrition), to test the hypothesis that adaptation of ewes to nutritional and environmental changes may alter placental efficiency and conceptus nutrient availability in the face of maternal nutrient restriction. Baggs ewes received 50 or 100% of the NRC nutrient requirements between d 28 and 78 of pregnancy. On d 78, maternal uterine arterial and fetal umbilical venous blood samples were obtained, and the ewes were euthanized. Amino acids and their metabolites (ammonia, urea, and polyamines) in plasma were analyzed using enzymatic and HPLC methods. The results showed that maternal plasma concentrations of 9 AA (Asp, Ile, Leu, Lys, Orn, Phe, Thr, Trp, and Val) as well as maternal and fetal plasma concentrations of ammonia and urea were reduced (P < 0.05) in nutrient-restricted compared with control-fed Baggs ewes. However, fetal plasma concentrations of all AA and polyamines did not differ (P = 0.842) between the 2 groups of ewes. Collectively, these findings suggest that Baggs ewes, by adapting to the harsh conditions and limited nutrition under which they were selected, were able to maintain fetal concentrations of AA in the face of a maternal nutrient restriction through augmenting placental efficiency.     

Effect of early gestational undernutrition on angiogenic factor expression and vascularity in the bovine placentome

The effect of early gestation maternal undernutrition followed by realimentation on placentomal vascular growth and angiogenic factor expression was determined in multiparous beef cows bred to the same bull. Cows gestating only female fetuses (n = 30) were fed in equal numbers to meet the NRC requirements (control) or were fed below the NRC requirements to lose BW (nutrient restricted; NR) from d 30 to 125 of gestation. After slaughter on d 125 of gestation, 10 control and 10 NR cows were necropsied. The remaining NR cows (n = 5) were then fed to achieve a BCS equal to their control group contemporaries (n = 5) by d 220 of gestation. All cows were fed the control diet from d 220 until 250 of gestation, when the remaining control and NR cows were slaughtered and necropsied. At necropsy, placentomes were fixed via perfusion of the caruncular and cotyledonary arteries to determine capillary vascular density. Cotyledonary (fetal placental) and caruncular (maternal placental) tissues also were snap-frozen in liquid nitrogen, and mRNA concentrations of vascular endothelial growth factor and its 2 specific receptors, fms-like tyrosine kinase and kinase insert domain containing receptor, as well as placental growth factor, were determined. There was no effect of diet or day of gestation on the percentage of proliferating caruncular cells. Although diet did not impact cotyledonary cellular proliferation, there was an increase (P < 0.05) in the percentage of proliferating cells on d 250 compared with d 125 of gestation. Nutrient restriction from d 30 to 125 increased (P < or = 0.10) placental mRNA concentrations of placental growth factor and fms-like tyrosine kinase; however, there was no alteration in vascularity. By d 250 of gestation, NR cows had increased (P < 0.05) caruncular capillary surface density and decreased (P < 0.05) cotyledonary capillary area density, capillary number density, and capillary surface density compared with control cows. Although nutrient restriction had little effect on placental vascularity by d 125, upon realimentation, alterations in vascularity became apparent by d 250 of gestation, suggesting a placental programming effect.     

Crop phenology modifies wheat responses to increased UV-B radiation

Ozone layer depletion increases the level of ultraviolet radiation reaching the earth's surface affecting both natural and agricultural ecosystems, especially in the Southern Hemisphere. Considering the harmful effects UV-B radiation has on plant growth the future productivity of wheat crops in Southern Chile could be challenged by both (i) the forthcoming level of UV-B increase and (ii) the sensitivity of this crop to higher UV-B radiation. In this study the effect of increased UV-B radiation at different phenophases on a spring wheat cultivar (Pandora) was investigated in two experiments at plant and crop levels under out-door conditions. The experiments consisted of controls, increased UV-B radiation at specific phenophases (from 3 leaf stage to booting 3L-Bo, and from booting to maturity Bo-PM), and increased UV-B radiation for the majority of the crop cycle (from 3 leaf stage to maturity). UV-B radiation was increased by Q panel UV-313 lamps set in plastic framed structures. Control plants were grown either without frames or below the same framed structures as those which received increased UV-B treatments. Phenology, above-ground biomass, grain yield, components, grain protein concentration, leaf area index (LAI), Fv/Fm and pigments were measured at booting and/or at harvest. Above-ground biomass and yield decreased by 11–19 and 12–20%, respectively, when UV-B radiation was increased at the 3L-Bo phase, while no effect was observed when irradiation was applied later in the crop cycle (Bo-PM). No additional UV-B effects to those observed at booting were detected in plants irradiated during the majority of the entire crop cycle (3L-PM). Biomass variation was strongly associated (r = 0.99; P < 0.01) with UV-B/PAR ratio in the sensitive treatments to UV-B increases (3 L-Bo) of both experiments. Flour protein was not affected by UV-B increases at any phenophase evaluated in this study. In both experiments, leaf green area and weight were negatively affected by increased UV-B radiation and no effect on specific leaf area (SLA) was found. Lower Fv/Fm, chlorophyll, carotenoid concentration and carotenoid:chlorophyll ratio were found at crop level (experiment 2) under higher UV-B in the 3L-Bo and 3L-PM treatments. The flavonoid concentration responded differently in the two experiments, probably due to the optimum responses these pigments had to expose UV-B doses.     

Identification of fatty acid esters of pectenotoxin-2 seco acid in blue mussels (Mytilus edulis) from Ireland

Pectenotoxins from marine dinoflagellates of the genus Dinophysis are rapidly hydrolyzed by many shellfish to give pectenotoxin-2 seco acid, which isomerizes to 7-epi-pectenotoxin-2 seco acid. Three series of fatty acid esters of pectenotoxin-2 seco acid (PTX-2 seco acid) and 7-epi-PTX-2 seco acid were detected by LC-MS analysis of extracts from blue mussels (Mytilus edulis) from Ireland. The locations of the fatty acid ester linkages were identified by a combination of LC-MSn in positive- and negative-ion modes, LC-MS analysis of the products from reaction of the esters with sodium periodate, and NMR analysis of purified samples of the two most abundant ester derivatives. The 37-O-acyl esters of PTX-2 seco acid were the most abundant, followed by the corresponding 11-O-acyl esters, accompanied by low levels of the 33-O-acyl esters. The most abundant fatty acid esters in the fractionated sample were, in order, the 16:0, 22:6, 14:0, 16:1, 18:4, and 20:5 fatty acids, although a wide array of other PTX-2 seco acid fatty acid esters were also present at low levels.     

Production and isolation of azaspiracid-1 and -2 from Azadinium spinosum culture in pilot scale photobioreactors

Azaspiracid (AZA) poisoning has been reported following consumption of contaminated shellfish, and is of human health concern. Hence, it is important to have sustainable amounts of the causative toxins available for toxicological studies and for instrument calibration in monitoring programs, without having to rely on natural toxin events. Continuous pilot scale culturing was carried out to evaluate the feasibility of AZA production using Azadinium spinosum cultures. Algae were harvested using tangential flow filtration or continuous centrifugation. AZAs were extracted using solid phase extraction (SPE) procedures, and subsequently purified. When coupling two stirred photobioreactors in series, cell concentrations reached 190,000 and 210,000 cell · mL(-1) at steady state in bioreactors 1 and 2, respectively. The AZA cell quota decreased as the dilution rate increased from 0.15 to 0.3 day(-1), with optimum toxin production at 0.25 day(-1). After optimization, SPE procedures allowed for the recovery of 79 ± 9% of AZAs. The preparative isolation procedure previously developed for shellfish was optimized for algal extracts, such that only four steps were necessary to obtain purified AZA1 and -2. A purification efficiency of more than 70% was achieved, and isolation from 1200 L of culture yielded 9.3 mg of AZA1 and 2.2 mg of AZA2 of >95% purity. This work demonstrated the feasibility of sustainably producing AZA1 and -2 from A. spinosum cultures.     

Morphology,tannin concentration and forage value of 15 Swiss accessions of sainfoin (Onobrychis viciifolia Scop.) as influenced by harvest time and cultivation site

Fifteen accessions of sainfoin (Onobrychis viciifolia Scop.) were characterized for morphological and phenological traits at Reckenholz in the Swiss lowlands (Experiment 1). The effects of accession, harvest time and site on dry‐matter yield, condensed tannin (CT) concentration and forage value (Experiment 2) were determined at three sites in Switzerland varying in altitude from 440 to 559 m. Three to four harvests were taken in the first year after establishment (second year of stand) with harvests 1 and 2 chemically analysed. From the characterization in Experiment 1, a clear grouping of single flowering (Communis) and multiple flowering (Bifera) accessions emerged. Additionally, within the Communis accessions, distinct groupings were identified (historical landraces and newly collected ecotypes) based on morphological characteristics. Experiment 2 showed that Communis and Bifera accessions had a similar chemical composition in the first harvest. In the second harvest, Communis accessions were higher in crude protein and CT and lower in neutral and acid detergent fibre concentrations than Bifera accessions. Total dry‐matter yields were higher for Bifera accessions. Among the Communis accessions, ecotypes had consistently higher CT concentrations than landraces. In vitro organic matter digestibility did not significantly differ among accessions. There were clear effects of harvest time and site for most variables, with clear harvest time × sainfoin group interaction but no site × sainfoin group interactions. The results clearly demonstrate that historical landraces and newly collected ecotypes expand the range of available genetic variation for sainfoin breeding.     

Effects of constant rate infusions of dexmedetomidine or MK-467 on the minimum alveolar concentration of sevoflurane in dogs

Objective

To determine the effects of low and high dose infusions of dexmedetomidine and a peripheral α₂-adrenoceptor antagonist, MK-467, on sevoflurane minimum alveolar concentration (MAC) in dogs.

Improved isolation procedure for azaspiracids from shellfish, structural elucidation of azaspiracid-6, and stability studies

Azaspiracids are a group of lipophilic polyether toxins produced by the small dinoflagellate Azadinium spinosum. They may accumulate in shellfish and can result in illnesses when consumed by humans. Research into analytical methods, chemistry, metabolism, and toxicology of azaspiracids has been severely constrained by the scarcity of high-purity azaspiracids. Consequently, since their discovery in 1995, considerable efforts have been made to develop methods for the isolation of azaspiracids in sufficient amounts and purities for toxicological studies, in addition to the preparation of standard reference materials. A seven-step procedure was improved for the isolation of azaspiracids-1-3 (1, 2, and 3) increasing recoveries 2-fold as compared to previous methods and leading to isolation of sufficiently purified azaspiracid-6 (6) for structural determination by NMR spectroscopy. The procedure, which involved a series of partitioning and column chromatography steps, was performed on 500 g of Mytilus edulis hepatopancreas tissue containing ~14 mg of 1. Overall yields of 1 (52%), 2 (43%), 3 (43%), and 6 (38%) were good, and purities were confirmed by NMR spectroscopy. The structure of 6 was determined by one- and two-dimensional NMR spectroscopy and mass spectrometry. The stability of 6 relative to 1 was also assessed in three solvents in a short-term study that demonstrated the greatest stability in aqueous acetonitrile.