Mycotoxin ingestion during late gestation alters placentome structure, cotyledon transcriptome, and fetal development in pregnant sheep.

Ergot alkaloids, a class of mycotoxins, induce vasoconstriction when consumed by animals and humans. Pregnant ewes (n = 16; 81.2 kg ± 7.7) were assigned fed endophyte-infected tall fescue seed (E+; 4.14 µg ergovaline + ergovalinine/g seed) or a control diet (CON; 0 µg ergovaline + ergovalinine) for increasing duration during late gestation (from gd86 to gd110 or gd133) to examine changes in placentome morphology and mRNA transcriptome, and fetal development. Exposure to E+ fescue reduced serum prolactin concentrations at gd110 and gd133 demonstrating treatment efficacy. For control ewes, cotyledon and total placentome weights decreased with advancing gestation due to remodeling of placental tissues; however, cotyledon and placentome weight did not change with advancing gestation in E+ fed ewes. Fetal brain sparing was evident in E+ exposed fetuses at gd110 and gd133 compared to CON, which demonstrates asymmetrical growth and intrauterine growth restriction. Mycotoxin exposure (E+) resulted in differential expression of 22 genes in the cotyledon tissue at gd110 but only one gene at gd133 compared to CON. These results suggest that the response to mycotoxin exposure in the pregnant sheep model has an immediate impact on placental remodeling and fetal development that persists throughout the duration of the exposure period.

Phytoextraction by harvesting dead leaves: cadmium accumulation associated with the leaf senescence in Festuca arundinacea Schreb.

Phytoextraction strategy by harvesting dead leaves provides continuous phytoremediation and a great saving in disposal cost of hazardous plant residues. This strategy is entirely dependent upon the amount of cadmium (Cd) accumulated in dead leaves. However, it is unknown that whether the leaf Cd accumulation is associated with its senescence and how to regulate its Cd accumulation. This study showed that Cd was preferentially and consistently distributed to and accumulated in the senescent leaves with the new leaf emergence and the old leaf dieback under 75 µM of Cd stress in tall fescue (Festuca arundinacea Schreb.). Individual leaf monitoring from its emergence to senescence showed that Cd concentration increased exponentially with the leaf life cycle, while leaf biomass decreased gradually after 14 days of leaf emergence. The total amount of Cd accumulated in the leaf showed an exponential increase during leaf senescence, regardless of the leaf biomass loss. Our results demonstrated that leaf Cd accumulation was significantly associated with its senescence and the highest Cd accumulated in dead leaves could be contributed from the continuous Cd input during the leaf senescent process, indicating that further regulatory studies should be focused on the leaf senescence process to achieve higher Cd accumulation and phytoextraction efficiency by harvesting dead leaves.

The effect of ergot alkaloid exposure during gestation on the microscopic morphology and vasculature of the ovine placenta.

Ergot alkaloids, a class of mycotoxins associated with ergotism, act as agonists on serotonin (5HT) receptors, specifically 5HT2a, which mediate smooth muscle contraction and vasoconstriction. The objective of this study was to examine the impact of ergot alkaloid exposure during mid and late gestation on microscopic placental structure and vascular development. Ewes were fed endophyte-infected tall fescue seed containing ergot alkaloids (E+/E+, 1.77 mg ewe-1 d-1) or endophyte-free tall fescue seed (E-/E-, 0 mg ergot alkaloids) during both mid (d 35 to d 85) and late gestation (d 86 to d 133). On d 133 of gestation, a terminal surgery was performed and two placentomes of the type B morphology were collected for microscopic analyses. Amorphous connective tissue regions were larger (p < 0.0001) and more numerous (p = 0.025) in the placentome of ergot alkaloid exposed ewes. Staining showed no difference (p = 0.83) in the number of vessels present, but luminal area of maternal vasculature was 117% greater (p < 0.0001) in ergot alkaloid exposed ewes. Results showed that exposure to ergot alkaloids during gestation slowed maturation of the fetal villi as indicated by greater amorphous connective tissue regions, and altered size and shape of blood vessels to counteract reductions in blood flow caused by vasoconstriction.

Antifeedant Effects and Repellent Activity of Loline Alkaloids from Endophyte-Infected Tall Fescue against Horn Flies, Haematobia irritans (Diptera: Muscidae).

Haematobia irritans is an obligate bloodsucking ectoparasite of cattle and is the global major pest of livestock production. Currently, H. irritans management is largely dependent upon broad-spectrum pesticides, which lately has led to the development of insecticide resistance. Thus, alternative control methods are necessary. Endophyte-infected grasses have been studied as an alternative due to their capability to biosynthesize alkaloids associated with anti-insect activities. Thus, the main aim of this study was to evaluate the antifeedant and repellent activity of lolines obtained from endophyte-infected tall fescue against H. irritans adults in laboratory conditions. The alkaloid extract (ALKE) was obtained by acid-base extraction. N-formyl loline (NFL) and N-acetyl loline (NAL) were isolated by preparative thin layer chromatography (pTLC) and column chromatography (CC), and the loline was prepared by acid hydrolysis of a NFL/NAL mixture. Loline identification was performed by gas chromatography coupled to mass spectrometry (GC/MS). Feeding behavior was evaluated by a non-choice test, and olfactory response was evaluated using a Y-tube olfactometer. Accordingly, all samples showed antifeedant activities. NFL was the most antifeedant compound at 0.5 µg/µL and 1.0 µg/µL, and it was statistically equal to NAL but different to loline; however, NAL was not statistically different to loline. NFL and NAL at 0.25 µg/µL were more active than loline. All samples except loline exhibited spatial repellency in the olfactometer. Thus, the little or non-adverse effects for cattle and beneficial activities of those lolines make them suitable candidates for horn fly management.

Mycelial biomass and concentration of loline alkaloids driven by complex population structure in Epichloë uncinata and meadow fescue (Schedonorus pratensis).

Many efforts have been made to select and isolate naturally occurring animal-friendly Epichloë strains for later reinfection into elite cultivars. Often this process involves large-scale screening of Epichloë-infected wild grass populations where strains are characterized and alkaloids measured. Here, we describe for the first time the use of genotyping-by-sequencing (GBS) on a collection of 217 Epichloë-infected grasses (7 S. arundinaceum, 4 L. perenne, and 206 S. pratensis). This genotyping strategy is cheaper than complete genome sequencing, is suitable for a large number of individuals, and, when applied to endophyte-infected grasses, conveniently genotypes both organisms. In total, 6273 single nucleotide polymorphisms (SNPs) in the endophyte data set and 38 323 SNPs in the host data set were obtained. Our findings reveal a composite structure with three distinct endophyte clusters unrelated to the three main S. pratensis gene pools that have most likely spread from different glacial refugia in Eurasia. All three gene pools can establish symbiosis with E. uncinata. A comparison of the endophyte clusters with microsatellite-based fingerprinting of the same samples allows a quick test to discriminate between these clusters using two simple sequence repeats (SSRs). Concentrations of loline alkaloids and mycelial biomass are correlated and differ significantly among the plant and endophyte subpopulations; one endophyte strain has higher levels of lolines than others, and one specific host genotype is particularly suitable to host E. uncinata. These findings pave the way for targeted artificial inoculations of specific host-endophyte combinations to boost loline production in the symbiota and for genome association studies with the aim of isolating genes involved in the compatibility between meadow fescue and E. uncinata.

Dispersive liquid-liquid microextraction as a preconcentration alternative to increase ETAAS sensitivity in the analysis of molybdenum in bovine meat and pasture samples.

A new procedure combining dispersive liquid-liquid microextraction and electrothermal atomic absorption spectrometry was proposed for the determination of molybdenum in plants and animal tissue. The optimized conditions for the microextraction procedure were: 150 µL of 0.3 mol L-1 potassium ethyl xanthate (pH 1.4) and 1000 µL of acetonitrile and 50 µL of carbon tetrachloride as dispersive and extractant solvent, respectively. A limit of detection of 0.03 µg kg-1 and a limit of quantification of 0.11 µg kg-1, as well as an enrichment factor of 200, were achieved employing this procedure. Standard reference materials were used for trueness and precision evaluation. No significant differences were found at the 95% confidence level between certified and obtained values, and the precision expressed as repeatability (sr %) was 4.1% for animal tissue and 4.7% for plant material. The method was successfully applied for the determination of Mo in bovine meat and pasture samples from different livestock establishments.

A novel phytoremediation method assisted by magnetized water to decontaminate soil Cd based on harvesting senescent and dead leaves of Festuca arundinacea.

It is more economical and feasible to harvest above-ground plant tissues, especially leaves, than to uproot the entire plant during phytoremediation. The major goal of this study is to estimate the promoting effect of an external magnetic field on soil Cadmium (Cd) remediation efficiency by harvesting the leaves of Festuca arundinacea, irrigated by normal and magnetized water. Changes in the amount of emerging, mature, senescent, and dead leaves, as well as Cd concentrations in the corresponding tissues were compared. It was found that F. arundinacea irrigated by either kind of water accumulated significantly more Cd in senescent and dead leaves than in other tissues. After irrigation with magnetized water, the biomass of senescent and dead leaves increased from 15.7 and 6.4% to 17.2 and 11.6%, respectively, and a significantly higher amount of Cd (˜23.6%) was redistributed into dead leaves compared with the control. Thus, significantly more Cd was removed by harvesting the senescent and dead leaves of F. arundinacea. These results indicate that magnetic fields can increase the phytoremediation efficiency of F. arundinacea for Cd, and simultaneously reduce the cost of harvested residue disposal.

Soil organic matter prevails over heavy metal pollution and vegetation as a factor shaping soil microbial communities at historical Zn-Pb mining sites.

This study examined the effects of soil heavy metals, macronutrients, texture and pH as well as plant species richness and composition on soil respiration, enzymatic activity, microbial biomass, metabolic quotient (qCO2) and arbuscular mycorrhizal fungi (AMF) at sites of historical Zn-Pb mining. The study was conducted both on a large scale (65 heaps scattered over the area of 750 km2) and on a small scale (25 plots along two 48 m transects extending from heaps to adjacent fallow fields). Total concentrations of metals exceeded 400 (Cd), 20,000 (Pb) and 80,000 (Zn) mg kg-1 at the most polluted sites. Although they decreased along the heap-fallow direction, they still remained above environmental standards in fallow soils. In contrast, some soluble metal forms increased with the increasing distance from heaps. Soil organic matter had the strongest positive effect on most microbial parameters. Total and/or available heavy metals exhibited significant negative effects on microbial biomass, enzymatic activity and AMF, and a positive effect on qCO2. Organic matter alleviated negative effects of heavy metals on microorganisms; they were not observed where the increase in the contamination was accompanied by the increase in organic matter content. Plant species richness affected positively enzymatic activity and mycorrhization level. Plant species composition possibly contributed to the formation of soil microbial communities, but its effect was entangled in that of heavy metals as plant communities changed along pollution gradients (from metal-tolerant grasslands dominated by Festuca ovina to calcareous grasslands and ruderal communities at less polluted sites).

Ruminal motility, reticuloruminal fill, and eating patterns in steers exposed to ergovaline.

Fescue toxicosis is problematic for growing steers, causing lower DMI and productivity when fed endophyte-infected (E+) tall fescue. A complete understanding of underlying mechanisms of how fescue toxicosis affects growing steers is lacking. Therefore, the overall objective of this multiexperiment study was to determine whether ruminally dosed ergovaline (ERV) affects rumen motility, rumen contents, and eating patterns. In Exp. 1, an 8-h period to assess ruminal motility began 4 h after feeding by monitoring pressure changes using a wireless system for 21 d. Eight ruminally cannulated steers (283 kg BW) were pair fed with alfalfa cubes (1.5 × NEm) and assigned to endophyte free (E-; 0 µg ERV/kg BW/d) or E+ treatment (20 µg ERV/kg BW/d). Overall, E+ steers had more frequent rumen contractions (Seed P = 0.05 and day of feeding P = 0.02). On days 7 to 9, both treatments showed lower frequencies and E- steers had greater amplitude of contractions (P < 0.001) that corresponded with decreased DMI. In Exp. 2, steers remained in pairs assigned in Exp. 1 (322 kg BW), but reversed seed treatments while increasing ERV levels (titrated 0, 5, 10, 15, and 20 µg ERV/kg BW/d over 57 d). There were no differences between E- and E+ for frequency (P = 0.137) or amplitude of contractions (P = 0.951), but increasing ERV dosage, decreased frequency (P = 0.018) and amplitude (P = 0.005), coinciding with lower DMI. In Exp. 3, 8 steers (589 kg) were pair fed and ruminally dosed 15 µg ERV/kg BW/d, and rumen motility data were collected for 21 d. E- steers showed higher amplitude and lower frequency of contractions than E+ steers with seed (P < 0.001), day (P < 0.001), and seed × day (P < 0.04) effects, but rumen fill was not different between E- and E+ (P > 0.29). Serum prolactin concentrations were lower in E+ steers in Exp. 1 to 3. Eating patterns of pair-fed E- and E+ steers were relatively slower in E+ than E- (Exp. 4) by measuring every 2 h across 24 h. Number of meals were higher in E+ than E- steers, but meal duration and meal size were not different between treatments. Rumen content (DM%) tended to be higher in E+ than in E- when steers were fed once a day (P = 0.07), but there was no difference for rumen content (DM%) when E- and E+ steers were fed 12 times a day (P = 0.13). These results suggest the changes in rumen fill associated with fescue toxicosis may be driven more by changes in feeding behavior and eating pattern rather than by changes in motility.

Assisted phytostabilization of soil from a former military area with mineral amendments.

Due to the presence of toxic pollutants, soils in former military areas need remedial actions with environmentally friendly methods. Greenhouse experiments were conducted to investigate the aided phytostabilization of multi-heavy metals (HMs), i.e. Cd, Cr, Cu, Ni, Pb, Zn, in post-military soil by Festuca rubra and three mineral amendments (diatomite, dolomite and halloysite). The amendments were applied at 0 and 3.0% to each pot filled with 5 kg of polluted soil. After seven weeks of the phytostabilization, selected soil properties, biomass yield of F. rubra and immobilization of HMs by their accumulation in plant and redistribution among individual fractions in soil were determined. In addition, ecotoxicology parameters of non-amended and amended soil were established using Phytotoxkit (Sinapsis alba) and Ostracodtoxkit (Heterocypris incongruens) tests. The addition of halloysite significantly increased F. rubra biomass. Diatomite significantly increased both the Cd, Cu, Pb and Cr concentrations in the roots and the pH of the soil. The application of halloysite significantly decreased the Cd and Zn contents of the soil after the completion of the experiment. Dolomite and halloysite were more effective in HM immobilization in soil by decreasing their content in an exchangeable fraction than diatomite. These soil amendments significantly differentiated the length of S. alba roots and had a positive effect on the development of H. incongruens.

Role of Festuca rubra and Festuca arundinacea in determinig the functional and genetic diversity of microorganisms and of the enzymatic activity in the soil polluted with diesel oil.

The objective of this study was to analyze the effect of two grass species, i.e. red fescue (Festuca rubra) and tall fescue (F. arundinacea), on the functional and genetic diversity of soil-dwelling microorganisms and on the enzymatic activity of soil not polluted and polluted with diesel oil. Grasses were examined for their effectiveness in accelerating degradation of PAHs introduced into soil with diesel oil. A growing experiment was conducted in Kick-Brauckman pots. The soil not polluted and polluted with diesel oil (7 cm3 kg-1 d.m.) was determined for the count of bacteria, colony development index, ecophysiological diversity index, functional diversity (using Biolog system), genetic diversity of bacteria (using NGS), enzymatic activity, and content of hydrocarbons. Study results demonstrated disturbed homeostasis of soil. The toxic effect of diesel oil on grasses alleviate with time since soil pollution. The yield of the first swath of red fescue decreased by 98% and that of tall fescue by 92%, whereas the yields of the second swath decreased by 82% and 89%, and these of the third swath by 50% and 47%, respectively. Diesel oil diminished also the functional and genetic diversity of bacteria. The use of grasses significantly decreased contents of C6-C12 (gasoline total), C12-C35 mineral oils, BTEX (volatile aromatic hydrocarbons), and PAHs in the soil, as well as enabled restoring the microbiological equilibrium in the soil, and increased functional and genetic diversity of bacteria. For this reason, both analyzed grass species, i.e. Festuca rubra and F. arundinacea, may be recommended for the remediation of soil polluted with diesel oil.

Effects of endophyte-infected tall fescue seed and protein supplementation on stocker steers: II. Adaptive and innate immune function.

Fescue toxicosis is a multifaceted syndrome common in cattle grazing endophyte-infected tall fescue that affects performance; however, little information is available pertaining to its effects on immunity. Recently, it has been shown that supplemental CP can improve performance in weaned steers postvaccination. Thus, the objective of this study was to evaluate the effect of supplemental CP on innate and adaptive immune responses in stocker steers chronically exposed to ergovaline. Angus steers (n = 12 pens; 3 steers/pen) were stratified by weight and assigned to a 2 × 2 factorial arrangement to examine crude protein levels of supplement (14% or 18%) and ergovaline exposure (0 or 185 µg ergovaline/kg BW/d via ground endophyte-free (EF) or endophyte-infected (EI) tall fescue seed, respectively) on immune response. Consumption of low to moderate concentration of ergovaline from EI tall fescue seed was sufficient to induce mild symptoms associated with fescue toxicosis. Blood samples were collected at day 0, 42, and 56 to evaluate infectious bovine rhinotracheitis (IBR) and bovine viral diarrhea virus (BVDV) type 1b titers following vaccine challenge. Additionally, serum cytokine concentrations were evaluated using Quantibody Bovine Cytokine Arrays on day 0, 28, and 42. Data were analyzed using PROC MIXED of SAS with repeated measures. Regardless of treatment, no differences were observed in IBR and BVDV-1b seroconversion following vaccine challenge (P > 0.05). Regardless of crude protein concentration, EI steers had greater concentrations of proinflammatory cytokines (TNF-α, IFN-γ, IL-1α), chemokines (CCL2, CCL4, MIG), anti-inflammatory cytokines (IL-2, -13, -15, -21), and various growth factors (FGF-1, IGF-1, VEGF-A) when compared to EF steers (P < 0.05). Furthermore, VEGF-A and IGF-1 concentrations were greater in EI-14 steers on day 28 compared to EI-18, EF-14, and EF-18 steers (P < 0.05), however, this difference was not observed on day 0 or 42 (P > 0.05). Based on these data, steers exposed to ergovaline have an increase in pro- and anti-inflammatory cytokines and supplemental CP had minimal impact to mitigate this response. However, in the current study, exposure to ergovaline had little to no effect on adaptive immunity and response to vaccination. Together, chronic exposure to ergovaline results in a hyperactive innate immune response, which may lead to an immuno-compromised animal.

Infection Rates and Alkaloid Patterns of Different Grass Species with Systemic Epichloë Endophytes.

Symbiotic Epichloë species are fungal endophytes of cool-season grasses that can produce alkaloids with toxicity to vertebrates and/or invertebrates. Monitoring infections and presence of alkaloids in grasses infected with Epichloë species can provide an estimate of possible intoxication risks for livestock. We sampled 3,046 individuals of 13 different grass species in three regions on 150 study sites in Germany. We determined infection rates and used PCR to identify Epichloë species diversity based on the presence of different alkaloid biosynthesis genes, then confirmed the possible chemotypes with high-performance liquid chromatography (HPLC)/ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and gas chromatography-mass spectrometry (GC-MS) measurements. Infections of Epichloë spp. were found in Festuca pratensis Huds. (81%), Festuca ovina L. aggregate (agg.) (73%), Lolium perenne L. (15%), Festuca rubra L. (15%) and Dactylis glomerata L. (8%). The other eight grass species did not appear to be infected. For the majority of Epichloë-infected L. perenne samples (98%), the alkaloids lolitrem B and peramine were present, but ergovaline was not detected, which was consistent with the genetic evaluation, as dmaW, the gene encoding the first step of the ergot alkaloid biosynthesis pathway, was absent. Epichloë uncinata in F. pratensis produced anti-insect loline compounds. The Epichloë spp. observed in the F. ovina agg. samples showed the greatest level of diversity, and different intermediates of the indole-diterpene pathway could be detected. Epichloë infection rates alone are insufficient to estimate intoxication risks for livestock, as other factors, like the ability of the endophyte to produce the alkaloids, also need to be assessed.IMPORTANCE Severe problems of livestock intoxication from Epichloë-infected forage grasses have been reported from New Zealand, Australia, and the United States, but much less frequently from Europe, and particularly not from Germany. Nevertheless, it is important to monitor infection rates and alkaloids of grasses with Epichloë fungi to estimate possible intoxication risks. Most studies focus on agricultural grass species like Lolium perenne and Festuca arundinacea, but other cool-season grass species can also be infected. We show that in Germany, infection rates and alkaloids differ between grass species and that some of the alkaloids can be toxic to livestock. Changes in grassland management due to changing climate, especially with a shift toward grasslands dominated with Epichloë-infected species such as Lolium perenne, may result in greater numbers of intoxicated livestock in the near future. We therefore suggest regular monitoring of grass species for infections and alkaloids and call for maintaining heterogenous grasslands for livestock.

Ergot alkaloid exposure during gestation alters: 3. Fetal growth, muscle fiber development, and miRNA transcriptome1.

The objective of this study was to assess how exposure to ergot alkaloids during 2 stages of gestation alters fetal growth, muscle fiber formation, and miRNA expression. Pregnant ewes (n = 36; BW = 83.26 ± 8.14 kg; 4/group; 9 groups) were used in a 2 × 2 factorial arrangement with 2 tall fescue seed treatments [endophyte-infected (E+) vs. endophyte-free (E-)] fed during 2 stages of gestation (MID, days 35 to 85 vs. LATE, days 86 to 133), which created 4 possible treatments (E-/E-, E+/E-, E-/E+, or E+/E+). Ewes were individually fed a total mixed ration containing E+ or E- fescue seed according to treatment assignment. Terminal surgeries were conducted on day 133 of gestation for the collection of fetal measurements and muscle samples. Data were analyzed as a 2 × 2 factorial with fescue treatment, stage of gestation, and 2-way interaction as fixed effects. Fetuses exposed to E+ seed during LATE gestation had reduced (P = 0.0020) fetal BW by 10% compared with E- fetuses; however, fetal body weight did not differ (P = 0.41) with E+ exposure during MID gestation. Fetuses from ewes fed E+ seed during MID and LATE gestation tended to have smaller (P = 0.058) kidney weights compared with E- fetuses. Liver weight was larger (P = 0.0069) in fetuses fed E- during LATE gestation compared with E+. Fetal brain weight did not differ by fescue treatment fed during MID (P = 0.36) or LATE (P = 0.40) gestation. The percentage of brain to empty body weight (EBW) was greater (P = 0.0048) in fetuses from ewes fed E+ fescue seed during LATE gestation, which is indicative of intrauterine growth restriction (IUGR). Primary muscle fiber number was lower (P = 0.0005) in semitendinosus (STN) of fetuses exposed to E+ during MID and/or LATE gestation compared with E-/E-. miRNA sequencing showed differential expression (P < 0.010) of 6 novel miRNAs including bta-miR-652_R+1, mdo-miR-22-3p, bta-miR-1277_R-1, ppy-miR-133a_L+1_1ss5TG, hsa-miR-129-1-3p, and ssc-miR-615 in fetal STN muscle. These miRNA are associated with glucose transport, insulin signaling, intracellular ATP, hypertension, or adipogenesis. This work supports the hypothesis that E+ tall fescue seed fed during late gestation reduces fetal weight and causes asymmetrical growth, which is indicative of IUGR. Changes in primary fiber number and miRNA of STN indicate that exposure to E+ fescue fed during MID and LATE gestation alters fetal muscle development that may affect postnatal muscle growth and meat quality.

The evaluation of resistance to Co 2+ of lawn plant at seedling stage and its concentration property at adult stage.

In order to evaluate the resistance of lawn plant to Co2+ at germination stage and discuss its concentration property at adult stage, four kinds of lawn plant which have some growth advantages in Co2+ polluted environment were selected as experimental materials in this research. They are tall fescue, timothy grass, inflorescences, and annual ryegrass. The results show that the evaluation of resistance to Co2+ of tall fescue's seed is the highest; the evaluation of resistance to Co2+ of annual ryegrass seed is the lowest. The low consistence Co2+ could improve the seed germination. With Co2+ concentration increase, the accumulation coefficient of four plants increased at first and decreased later; the accumulation coefficient of underground portion is higher than the accumulation coefficient of aboveground; with Co2+ concentration increase, the transfer coefficient of four plants have a remarkable decline. In these plants, the accumulation coefficient of tall fescue and annual ryegrass is bigger than other two plants. As the concentration of Co2+ treatment increased to 100 mg/kg, the aboveground enrichment of Co2+ in F. elata reached 75 mg/kg, followed by L. multiflorum (68.9 mg/kg), P. pratense (48.8 mg/kg), and D. glomerata (27.2 mg/kg).The highest underground enrichment of Co2+ in F. elata reached 836.46 mg/kg, in contrast to the lowest underground enrichment in D. glomerata, 264.67 mg/kg. It shows that fescue and annual ryegrass have a better enrichment property to Co2+ and have a better prospect for the treatment of cobalt-contaminated soil. This research could provide some scientific basis and main technical approach for the soil contaminated by Co2+.

Ergot alkaloid exposure during gestation alters. I. Maternal characteristics and placental development of pregnant ewes1.

Tall fescue [Lolium arundinaceum (Scheyreb.) Darbysh] is the primary cool season forage grass in the Southeastern United States. Most tall fescue contains an endophytic fungus (Epichloë coenophiala) that produces ergot alkaloids and upon ingestion induces fescue toxicosis. The objective of this study was to assess how exposure to endophyte-infected (E+; 1.77 mg hd-1 d-1 ergovaline and ergovalinine) or endophyte-free (E-; 0 mg hd-1 d-1 ergovaline and ergovalinine) tall fescue seed fed during 2 stages of gestation (MID, days 35-85/LATE, days 86-133) alters placental development. Thirty-six, fescue naïve Suffolk ewes were randomly assigned to 1 of 4 fescue treatments: E-/E-, E-/E+, E+/E-, or E+/E+. Ewes were individually fed the same amount of E+ or E- seed mixed into total mixed ration during MID and LATE gestation. Terminal surgeries were conducted on day 133 of gestation. Ewes fed E+ fescue seed had elevated (P < 0.001) ergot alkaloid excretion and reduced (P < 0.001) prolactin levels during the periods when fed E+ seed. Ewes switched on day 86 from E- to E+ seed had a 4% reduction (P = 0.005) in DMI during LATE gestation, which translated to a 2% reduction (P = 0.07) in DMI overall. Average daily gain was also reduced (P = 0.049) by 64% for E-/E+ ewes during LATE gestation and tended to be reduced (P = 0.06) by 33% overall. Ewes fed E+ seed during LATE gestation exhibited a 14% and 23% reduction in uterine (P = 0.03) and placentome (P = 0.004) weights, respectively. Caruncle weights were also reduced by 28% (P = 0.003) for E-/E+ ewes compared with E-/E- and E+/E-. Ewes fed E+ seed during both MID and LATE gestation exhibited a 32% reduction in cotyledon (P = 0.01) weights, whereas ewes fed E+ seed only during MID gestation (E+/E-) had improved (P = 0.01) cotyledon weights. The percentage of type A placentomes tended to be greater (P = 0.08) for E+/E+ ewes compared with other treatments. Other placentome types (B, C, or D) did not differ (P > 0.05). Total fetal weight per ewe was reduced (P = 0.01) for ewes fed E+ seed during LATE gestation compared with E-; however, feeding E+ seed during MID gestation did not alter (P = 0.70) total fetal weight per ewe. These results suggest that exposure to ergot alkaloids during LATE (days 86-133) gestation has the greatest impact on placental development by reducing uterine and placentome weights. This, in turn, reduced total fetal weight per ewe by 15% in ewes fed E+ seed during LATE gestation (E-/E+ and E+/E+).

Ergot alkaloid exposure during gestation alters: II. Uterine and umbilical artery vasoactivity1.

Previous research has shown that livestock exposed to ergot alkaloids results in decreased vasoactivity of gastrointestinal and peripheral vasculature. Little is known regarding the effect ergot alkaloid exposure during gestation may have on vasculature supporting the fetus. The objective of this study was to evaluate contractile responses of uterine and umbilical arteries collected from ewes consuming ergot alkaloids during gestation. On day 35 of gestation, 36 Suffolk ewes (78.24 ± 9.5 kg) were assigned to endophyte-infected (E+) or endophyte-free (E-) tall fescue seed treatments that were fed either throughout or switched on day 86 of gestation, creating four seed treatments E+E+, E+E-, E-E+, and E-E-. Ewes were fed E+ tall fescue seed to provide 1.77 mg of total ergovaline ⋅ hd-1 ⋅ d-1 with E- ewes receiving the same quantity of E- seed. Gestation was terminated on day 133, and sections of uterine artery and umbilical cord were surgically collected. Only collections from 28 ewes (n = 7/treatment) were of sufficient viability to proceed with the contractility experiments. Arteries were cleaned, sliced into 2-mm cross sections, and suspended in multi-myograph chambers containing 5 mL of continuously oxygenated Krebs-Henseleit buffer. Vessels were exposed to increasing concentrations (5 × 10-8 to 1 × 10-4 M) of norepinephrine, serotonin, ergotamine, and ergovaline (5 × 10-9 to 1 × 10-5M; extract of tall fescue seed) in 15-min intervals. Increasing concentrations of norepinephrine generated a contractile response by the uterine artery (P < 0.05), but no response in the umbilical artery. Increasing concentrations of serotonin resulted in negligible responses in uterine preparations, whereas umbilical artery preparations were responsive (P < 0.05) to serotonin. Ewes receiving E+E+ and E-E+ treatments had decreased vasoactivity in umbilical arteries to serotonin with a dextral shift in concentrations where the response curve initiated (P < 0.05). Interestingly, uterine arteries were not responsive to exposure to ergotamine or ergovaline, whereas umbilical arteries were responsive (P < 0.05). Umbilical arteries collected from ewes receiving E-E- and E+E- were more vasoactive to ergot alkaloids (P < 0.05) than other treatments. These findings indicate that maternal blood supply to the placenta appears protected from negative effects of ergot alkaloids; however, umbilical vasculature is not, and this could adversely influence fetal growth.

Effect of microbial inoculants on the fermentation characteristics, nutritive value, and in vitro digestibility of various forages.

This study was aimed to examine the characteristics of three isolated lactic acid bacteria (LAB) strains (HG24, M1, and TG1), and to evaluate their effects on fermentation characteristics, nutritive value, and in vitro digestibility in a variety of substrates. The experiments were conducted with Italian ryegrass (Lolium multiflorum Lam.), tall fescue (Festuca arundinacea Schred.), and oat (Avena sativa L.) in laboratory silo (1 L) for ensiling 30 days. All the treated silages had significantly (p < 0.05) higher in vitro dry matter digestibility (IVDMD), and lower undesirable microorganism populations compared with controls. In Italian ryegrass and oat silages, all the inoculants improved the fermentation quality, evidenced by distinctly (p < 0.05) higher LA contents and ratios of LA to acetic acid, and lower pH, butyric acid and ammonia nitrogen contents. Strains HG24 and commercial inoculant G (Lactobacillus plantarum MTD-1) performed better than strains M1 and TG1, indicated by obviously (p < 0.05) higher LA contents, LAB numbers, and IVDMD. In tall fescue silage, all the isolated LAB strains (HG24, M1 and TG1) markedly (p < 0.05) increased IVDMD compared with strain G. Strain HG24 is recommended as a good LAB inoculant for ensiling in various forages.

Forms of selenium in vitamin-mineral mixes differentially affect the expression of genes responsible for prolactin, ACTH, and α-MSH synthesis and mitochondrial dysfunction in pituitaries of steers grazing endophyte-infected tall fescue.

The goal of this study was to test the hypothesis that sodium selenite (inorganic Se, ISe), SEL-PLEX (organic forms of Se, OSe), vs. a 1:1 blend (MIX) of ISe and OSe in a basal vitamin-mineral (VM) mix would differentially alter pituitary transcriptome profiles in growing beef steers grazing an endophyte-infected tall fescue (E+) pasture. Predominately Angus steers (BW = 183 ± 34 kg) were randomly selected from fall-calving cows grazing E+ pasture and consuming VM mixes that contained 35 ppm Se as ISe, OSe, or MIX forms. Steers were weaned, depleted of Se for 98 d, and subjected to summer-long common grazing of a 10.1 ha E+ pasture containing 0.51 ppm ergot alkaloids. Steers were assigned (n = 8 per treatment) to the same Se-form treatments on which they were raised. Selenium treatments were administered by daily top-dressing 85 g of VM mix onto 0.23 kg soyhulls, using in-pasture Calan gates. As previously reported, serum prolactin was greater for MIX (52%) and OSe (59%) steers vs. ISe. Pituitaries were collected at slaughter and changes in global and selected mRNA expression patterns determined by microarray and real-time reverse transcription PCR analyses, respectively. The effects of Se treatment on relative gene expression were subjected to one-way ANOVA. The form of Se affected the expression of 542 annotated genes (P < 0.005). Integrated pathway analysis found a canonical pathway network between prolactin and pro-opiomelanocortin (POMC)/ACTH/α-melanocyte-stimulating hormone (α-MSH) synthesis-related proteins and that mitochondrial dysfunction was a top-affected canonical pathway. Targeted reverse transcription-PCR analysis found that the relative abundance of mRNA encoding prolactin and POMC/ACTH/α-MSH synthesis-related proteins was affected (P < 0.05) by the form of Se, as were (P ≤ 0.05) mitochondrial dysfunction-related proteins (CYB5A, FURIN, GPX4, and PSENEN). OSe steers appeared to have a greater prolactin synthesis capacity (more PRL mRNA) vs. ISe steers through decreased dopamine type two receptor signaling (more DRD2 mRNA), whereas MIX steers had a greater prolactin synthesis capacity (more PRL mRNA) and release potential by increasing thyrotropin-releasing hormone concentrations (less TRH receptor mRNA) than ISe steers. OSe steers also had a greater ACTH and α-MSH synthesis potential (more POMC, PCSK2, CPE, and PAM mRNA) than ISe steers. We conclude that form of Se in VM mixes altered expression of genes responsible for prolactin and POMC/ACTH/α-MSH synthesis, and mitochondrial function, in pituitaries of growing beef steers subjected to summer-long grazing an E+ pasture.

A novel phytoextraction strategy based on harvesting the dead leaves: Cadmium distribution and chelator regulations among leaves of tall fescue.

The treatment of large amount of hazardous plant residues from phytoextraction is costly and has been a burden for the society. This experiment was designed to evaluate the possibility of cadmium (Cd) phytoextraction by harvesting the dead leaves instead of the whole plant in tall fescue (Festuca arundinacea). Results showed that Cd was preferentially distributed in the senescent and dead leaves. EDTA, DTPA and EGTA enhanced Cd accumulations in the dead leaves which could be associated to the increase of the water-soluble inorganic Cd and Cd-organic acid complexes in shoots. The dead leaves were only 12.6-16.3% of the total shoot biomass but accumulated 73.4-87.2% of the total shoot Cd. The results indicate that a novel strategy of Cd phytoextraction based on harvesting the dead leaves is feasible to save the high treatment cost of hazardous plant residues while maintaining the acceptable phytoextraction efficiency.