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Characterization of Dac g 4, a major basic allergen from Dactylis glomerata pollen.

Monoclonal antibodies were produced against Dac g 4, a purified major basic allergen from Dactylis glomerata pollen. Their ability to be used for immunopurification of Dac g 4 was studied on a BIAcore apparatus (Pharmacia). The allergen was purified by affinity chromatography with one monoclonal antibody. Its precise molecular mass, 59,185 +/- 30 d, was determined by mass spectrometry. Its isoelectric point is 10.4. Sodium dodecylsulfate-polyacrylamide gel electrophoresis and immunoblotting showed that Dac g 4-related proteins of similar molecular mass were detected in the majority of allergenic grass pollen species. By double-site ELISAs, we have estimated that Dac g 4 represents about 6% of the total proteins from a water-soluble extract. One monoclonal antibody (mAb H) recognized a 60 kd cross-reactive protein in other grass pollens, though none in any of the tree or weed pollens tested. Inhibition studies of IgE antibody binding to Dac g 4 with pollen extracts confirmed the presence of cross-reactive allergens in Secale cereale, Lolium perenne, Festuca elatior, Holcus lanatus, Bromus arvensis, Poa pratense, Hordeum sativum, and Phleum pratense.

Allergens↗

How far can we simplify in vitro diagnostics for grass pollen allergy?: A study with 17 whole pollen extracts and purified natural and recombinant major allergens.

BACKGROUND: Current diagnostics for grass pollen allergy are composed of mixtures of pollen of different grass species. Their complex composition hampers accurate standardization. OBJECTIVE: The aim of the study was to investigate whether mixtures of grass pollen extracts can be replaced by a single pollen species and whether a single pollen species can be replaced by a limited number of purified natural or recombinant major allergens. METHODS: Sera (n = 800) were selected on the basis of a general suspicion for inhalant allergy and tested in a RAST for IgE reactivity with pollen from 17 different grass species. Cross-reactivity of IgE responses was studied by means of RAST inhibition. Sera with positive test results for grass pollen were tested in a RAST for natural Lol p 1 and Lol p 5 and recombinant Phl p 1 and Phl p 5. RESULTS: Specific IgE antibodies against one or more of the 17 pollen species were detected in 209 of 800 sera (26.1%). The highest responses were observed against Poa pratensis followed by Festuca rubra, Phleum pratense, and Dactylis glomerata. IgE responses were clearly lower (approximately by a factor of 5) against only three species (Phragmites communis, Cynodon dactylon, and Zea mays). With the exception of a few low-responder sera, no sera were found to have negative test results to the high responder species and positive results to any of the other species. Sera with positive test results for grass pollen (n = 154) were tested with purified Lol p 1 and Lol p 5. IgE anti-Lol p 1 and Lol p 5 accounted for an average of 81% +/- 7% of total anti-grass pollen IgE. For 14 sera (all with low anti-grass pollen IgE titers), a RAST with purified allergens resulted in a false-negative diagnosis for grass pollen allergy. With recombinant Phl p 1 and Phl p 5, the mean IgE reactivity was 57% +/- 6% of the anti-grass pollen IgE response (n = 141), with 13 false-negative results. CONCLUSION: One grass species is sufficient for in vitro diagnosis of grass pollen allergy. With purified natural Lol p 1 and Lol p 5, greater than 90% of grass-positive sera is detected. Around 80% of the IgE response to grass pollen is directed to these major allergens. Recombinant allergens, produced in Escherichia coli, did not equal the IgE-binding capacity of their natural counterparts.

Allergens↗

Impact of consuming tall fescue seeds infected with the endophytic fungus, Neotyphodium coenophialum, on reproduction of chickens.

Most of the 14 million hectares of pastures of tall fescue (Festuca arundinacea) planted in the United States are infected with the endophytic fungus, Neotyphodium coenophialum. I examined whether the consumption of fescue seed infected with this fungus had an adverse impact on reproduction in birds. Chickens were used as the model; they were maintained on a diet of chick starter mixed in equal amounts with either infected fescue seed (fungus-fed chickens) or uninfected fescue seed (control chickens). Consumption of infected fescue seed slowed the growth rates of female chicks but not males. Fungus-fed hens gained weight more slowly than control hens and also produced fewer eggs. Egg weight and fertility were similar in both groups of hens. Fungus-fed hens were more likely than control hens to produce eggs which lacked a shell. Naive hens exhibited an aversion for infected fescue seed when the seed was first presented to them, but the aversion disappeared after the hens were placed on a diet which included fescue seed for 3 or 6 months. Hens did not discriminate between feed containing infected seed and feed containing uninfected seed.

Animal Feed↗

Involvement of antioxidants and lipid peroxidation in the adaptation of two cool-season grasses to localized drought stress.

In natural environments, drought often occurs in surface soil while water is available for plant uptake deeper in the soil profile. The objective of the study was to examine the involvement of antioxidant metabolism and lipid peroxidation in the responses of two cool-season grasses to surface soil drying. Kentucky bluegrass (Poa pratensis L) and tall fescue (Festuca arundinacea Schreb.) were grown in split tubes, consisting of two sections (each 10 cm in diameter and 20 cm long). Grasses were subjected to three soil moisture regimes: (a) well-watered control: whole soil profile was watered; (b) surface drying: surface 20 cm of soil was dried by withholding irrigation and the lower 20 cm of soil was watered; (c) full drying: whole soil profile was dried. Surface drying had no effects on relative water content (RWC) and chlorophyll content (Chl) for both grasses and only slightly reduced shoot growth for tall fescue. Superoxide dismutase (SOD) activity increased, while catalase (CAT) and peroxidase (POD) activities remained unchanged during most periods of surface drying. Malondialdehyde (MDA) content was unaffected by surface drying for tall fescue, but increased initially and then decreased to the control level for Kentucky bluegrass. Under full drying, RWC, Chl content, and shoot dry weight decreased, but MDA content increased in both grasses; SOD and POD activities initially increased transiently and then decreased; CAT remained unchanged for 25 days and then decreased. These results suggested that both Kentucky bluegrass and tall fescue were capable of surviving surface soil drying. This capability could be related to increases in antioxidant activities, particularly SOD and CAT. However, full drying suppressed antioxidant activities and induced lipid peroxidation.

Journal Article↗

Oral and parenteral vaccination of mice with protein-ergotamine conjugates and evaluation of protection against fescue toxicosis.

Acremonium coenophialum produces ergopeptide alkaloids in tall fescue (Festuca arundinacea Schreb.). These ergot alkaloids decrease serum alkaline phosphatase (ALP) activity, serum cholesterol and prolactin concentrations, as well as average daily gains (ADG) in cattle. The objective of this study was to evaluate the protection of anti-ergotamine antibodies induced by either oral or parenteral vaccination with protein-ergotamine conjugates or passive vaccination with anti-ergovaline, monoclonal antibodies in a murine model of fescue toxicosis. Ergotamine (EG) was conjugated to bovine serum albumin (BSA) and cholera toxin subunit B (CTB) by the Mannich reaction. Mice were blocked based on weight and randomly allocated into five groups of 10 mice each. Treatment groups were as follows: (1) group vaccinated intraperitoneally (ip) with a BSA-EG conjugate and fed an endophyte-infected (EI) fescue diet (BSA-EG group); (2) group orally vaccinated with a CTB-EG conjugate mixed with free cholera toxin (CT) and fed an EI fescue diet (CTB-EG group); (3) nonvaccinated group fed an EI fescue diet (EI group); (4) group passively vaccinated with anti-ergovaline, monoclonal antibodies and fed an EI fescue diet (MoAB group); and (5) nonvaccinated group fed an endophyte-free (EF) fescue diet (EF group). The EI diet contained 1.5 ppm of Ergovaline (EV), whereas no EV was detected in the EF diet.Respective diets were similar upon nutritional analysis. Unvaccinated mice in the EI group exhibited features of fescue toxicosis as indicated by decreased serum ALP activity and cholesterol, and decreased weight gain as compared to mice in the EF group. Antibodies against EG and EV were present in sera of mice in the BSA-EG and MoAB groups, respectively. Mice orally vaccinated with the CTB-EG conjugate developed secretory IgA (sIgA) antibodies and short-lived, systemic IgG responses against EG. Weight gains were increased in the BSA-EG and CTB-EG groups and tended to be increased in the MoAB group vs. the unvaccinated EI group. Serum ALP activity was decreased in the BSA-EG and MoAB groups as compared to the EF group. Serum ALP activity was further decreased in the BSA-EG vaccinated group as compared to the EI group. Cholesterol concentrations were decreased in the EI, BSA-EG and MoAB groups as compared to the EF group. Prolactin concentrations were similar in all groups.

Acremonium↗

Filovirus-like particles detected in the leafhopper Psammotettix alienus.

Filamentous particles were detected by negative contrast electron microscopy of extracts from the leafhopper Psammotettix alienus (Dahlbom) reared on healthy Festuca gigantea plants. The particles were straight, slightly curved or flexuous, sometimes with the one end curled into a ring with an outer diameter of about 200 nm. Length distribution of 280 particles showed a minor and a major group with median length of about 600 and 1,100 nm, respectively. Projections, 8-10 nm long and about 10 nm apart, were evenly distributed on the surface. The diameter of particles, including projections, was 55-70 nm. Partly disintegrated revealed an internal structure about 30 nm in diameter and with cross-striation with a periodicity of 5-5.5 nm. In some particles, a central canal, 5-10 nm in diameter, could be seen at one end. Ultramicrotomy of leafhopper heads showed that some cells contained intracytoplasmic clusters of particles together with filamentous structures. The particles described in this paper resemble virions in the virus family Filoviridae, but can be distinguished by having a smaller particle diameter. The name Taastrup virus is suggested for the putative virus from Psammotettix alienus, according to the place it was first detected.

Animals↗

Behaviour and transport of radionuclides in soil and vegetation of a sand dune ecosystem.

A sand dune ecosystem in the vicinity of the British Nuclear Fuels reprocessing plant at Sellafield, Cumbria, UK was used to examine the spatial, temporal and depth distributions of 134Cs, 137Cs, 238Pu, 239 + 240Pu and 241Am in soil and in two species of vegetation (Festuca rubra, Ammophila arenaria). Core samples showed evidence of the accumulation of radionuclides derived mainly from sea-to-land transfer. Accumulated deposits of radioactivity (0-0.1 m) lie within the range: 1.1-3.4 Bq kg-1 (134Cs), 260-440 Bq kg-1 (137Cs), 31-40 Bq kg-1 (238Pu), 150-215 Bq kg-1 (239 + 240Pu) and 190-240 Bq kg-1 (241Am). Soil profiles showed greater activity concentrations in their deeper regions and this is attributed to leaching of radionuclides in percolating drainage water accentuated by the coarse texture, low organic matter and clay mineral content of coastal sands. Radionuclide activity concentrations in F. rubra and A. arenaria were similar, in the ranges 20-70 Bq kg-1 (137Cs), 1-5 Bq kg-1 (238Pu), 10-30 Bq kg-1 (239 + 240Pu) and 10-65 Bq kg-1 (241Am). Clear temporal and spatial variations were observed in both species of vegetation, reflecting the weather conditions antecedent to the sampling period and the influence of sea-to-land transfer. Concentration ratios (vegetation:soil) for activity concentrations in the two species were similar, in the ranges: 0.05-0.14 (137Cs), 0.025-0.097 (238Pu), 0.022-0.057 (239 + 240Pu) and 0.025-0.212 (241Am).

Americium↗

The influence of nitrogen deposition, competition and desiccation on growth and regeneration of Racomitrium lanuginosum (Hedw.) brid.

Racomitrium lanuginosum shoot growth was studied under the combined effects of N deposition (0, 20, 40 and 60 kg N ha(-1) year(-1)). competition with Festuca ovina, and a drought pre-treatment. Moss regeneration from shoot fragments was also investigated. Growth was initially stimulated at the 60 kg N level. However, after 6 months, growth was lower in all N treatments than in the 0 kg N control. Reductions in shoot growth first became apparent in the pre-desiccated moss, while moss shoots grew longer when surrounded by a F. ovina canopy. Optimum regeneration occurred at 20-40 kg N on bare soil, and at 0-20 kg N under a F. ovina canopy. These results suggest that current N deposition in upland Wales is already detrimental to growth of this species, and to regeneration under certain conditions. This species may be affected under predicted climatic scenarios of increased summer drought in Britain.

Air Pollutants↗

Reclamation of a bare industrial area contaminated by non-ferrous metals: physico-chemical and biological evaluation of the durability of soil treatment and revegetation.

In 1990, 3 ha of a highly metal polluted acid sandy soil at the site of a former pyrometallurgical zinc smelter was treated with a combination of beringite and compost; beringite is a substance that has a strong metal immobilization capacity. After soil treatment and sowing of a mixture of metal-tolerant Agrostis capillaris and Festuca rubra, a healthy vegetation cover developed. Five years later, an evaluation was made of soil physico-chemical parameters, potential phytotoxicity, floristic and fungal diversity and mycorrhizal infection of the plant community. Phytotoxicity was shown to be maintained at the low level observed immediately after soil treatment. The water-extractable metal fraction of the treated soil was up to 70 times lower compared to the non-treated soil. The vegetation was still healthy and regenerating by vegetative means and by seed. Diversity of higher plant species and saprophytic fungi was extremely low in the untreated area due to the high soil toxicity and the absence of metal tolerant ecotypes of plants and fungi. On the treated soil, in contrast, the species richness of higher plants was much higher; several perennial forbs which are not noted as metal tolerant had colonized the revegetated area. Most of these species belong to mycotrophic families so that the presence of a mycorrhizal network in the soil promotes their establishment. The ubiquity of the mycorrhizal fungi in the roots showed that a functioning ecosystem was establishing. In non-treated soil, the mycorrhizal infection rates of the roots were consistently lower during the whole growing season.

Journal Article↗

A comparison of stable caesium uptake by six grass species of contrasting growth strategy.

Six plant species in the family Gramineae were used to investigate the relationship between Cs uptake, nutrient regime and plant growth strategy sensu Grime (1979: Plant Growth Strategies and Vegetation Processes, John Wiley). The roots of 66 day old Elymus repens (L.) Gould., Bromus sterilis L., Agrostis stolonifera L., Anthoxanthum odoratum L., Festuca ovina L. and Nardus stricta L. plants grown in acid-washed sand at high and low nutrient levels were exposed to a 96 h pulse of stable Cs at 0.05 mM, 0.15 mM, 0.3 mM, 1.0 mM and 3.0 mM concentrations. Different nutrient regimes induced large differences in dry wt in E. repens, B. sterilis and A. stolonifera plants but only small differences in N. stricta and F. ovina plants. At high nutrient concentrations, A. stolonifera, A. odoratum, F. ovina and N. stricta shoots showed significantly greater increases in internal Cs concentration with rising external Cs concentrations than did E. repens and B. sterilis shoots. The relationship between increases in shoot and external Cs concentrations was statistically indistinguishable between species in plants grown at the low nutrient concentration. These patterns of Cs uptake ensured that with long-term high K concentrations the more competitive plants (E. repens and B. sterilis) accumulated higher concentrations of Cs from low external concentrations than did non-competitive plants or competitive plants grown at low nutrient levels. It is suggested that the relationship between plant growth strategy sensu Grime (1979) and Cs accumulation patterns may help to explain the different concentrations to which species accumulate radiocaesium from the soil.

Journal Article↗

High-performance liquid chromatographic determination of 4-methylimidazole in sheep plasma and in ammoniated tall fescue hay.

A method for 4-methylimidazole (4MI) extraction and quantitation in body fluids and forage samples was developed. The procedures involve ion-pair extraction of the compound with the quantitation done by ion-pair liquid chromatography. The results indicate that this high-performance liquid chromatographic method is sensitive, reproducible and more rapid than others that have been previously used. The mean recovery of 4MI from plasma and tall fescue (Festuca arundinacea) hay samples were above 95 and 85%, respectively. The versatility of the procedure makes it suitable for the determination of 4MI in body fluids and in forage samples.

Ammonia↗

Antithrombin activity of medicinal plants of the Azores.

A chromogenic bioassay was utilized to determine the antithrombin activity of methylene chloride and methanol extracts prepared from 50 plants of Azores. Extracts of the six plants Hedychium gardneranum, Tropaeolum majus, Gunnera tinctoria, Hedera helix, Festuca jubata and Laurus azorica demonstrated activity of 78% or higher in this bioassay system.

Azores↗

Poaceae in the natural diet of the snail Helix aspersa Müller (Gastropoda, Pulmonata).

The natural diet of Helix aspersa was studied in two populations by analysing faeces. Picris echioides, Carduus tenuifloris, Urtica dioica, Galium molugo (Dicotyledons) and Poaceae (Monocotyledons) were the principal resources of the studied populations. The species of Poaceae ingested by Helix aspersa in July were determined by analysing the phytoliths present in the faeces. Festuca rubra, Dactylis glomerata and Bromus hordeaceus seemed to be attractive for the snails whereas Elytrigia repens was rejected. These results were convergent with the ingestion rates of Poaceae in the laboratory but assimilation efficiencies did not explain these choices. Poaceae with the highest energetic values were not preferred to others. The texture, the mineral and organic contents of the Poaceae may be responsible for the snails feeding choices. The importance of Poaceae for this species is discussed.

Animals↗

Effects of the fermentation product of herbs by lactic acid bacteria against phytopathogenic filamentous fungi and on the growth of host plants.

The fermentation product of herbs by lactic acid bacteria (FHL) was assayed for antifungal activities against Rosellinia necatrix, Helicobasidium mompa, Fusarium oxysporum, Pythium graminicola and Pyricularia oryzae. FHL completely inhibited the growth of R. necatrix, H. mompa, P. graminicola and P. oryzae, and reduced the growth of F. oxysporum by 35%. When the seeds of Medicago sativa L. (alfalfa), Asparagus officinalis L. (asparagus), Brassica campestris L. (komatsuna), Oryza sativa L. (rice), Spinacia oleracea L. (spinach), Festuca arundinacea Schreb. (tall fescue), and Lycopersicum esculentum Mill. (tomato) were put on plates containing 0.69 mg/ml FHL, their germination rates did not decrease. The root elongation of A. officinalis, B. campestris, O. sativa, and L. esculentum seedlings was suppressed on plates containing 6.92 mg/ml FHL, but the root elongation of M. sativa was not suppressed on the 6.92 mg/ml FHL plate. When FHL was diluted to less than 1.73 mg/ml, the diluted FHL solution did not suppress the germination of B. campestris seeds, but it suppressed the root elongation of B. campestris seedlings. An FHL concentration higher than 0.35 mg/ml hastened the growth of seedlings of B. campestris in the presence of a chemical fertilizer but delayed the growth of these seedlings in the absence of the chemical fertilizer, suggesting that inorganic elements could affect the efficiency of FHL.

Journal Article↗

Fungal endophyte N-acetylglucosaminidase expression in the infected host grass.

Fungal endophytes of the genera Neotyphodium and Epichlolë are important mutualistic symbionts and pathogens of many cool-season grass species. Here we report the characterization of a secreted N-acetylglucosaminidase from the Neotyphodium sp. endophyte that infects the grass Poa ampla. The enzyme was expressed at low levels within the host, and activity could be detected in the apoplastic protein fraction. Low-level expression could also be detected in endophyte-infected perennial ryegrass (Lolium perenne), Chewings fescue (Festuca rubra subsp. fallax), and tall fescue (L. arundinaceum). The enzyme may function in the recycling of chitin oligomers generated from turnover of the fungal cell wall. This is the first report of a secreted N-acetylglucosaminidase expressed by an endophytic fungus in the infected host plant.

Acetylglucosaminidase↗

Mobility and dissipation of ethofumesate and halofenozide in turfgrass and bare soil.

The effect of turfgrass cover on the leaching and dissipation of ethofumesate and halofenozide was studied. Sampling cylinders (20 cm diam. x 30 cm long) were placed vertically in plots of creeping bentgrass (Agrostis palustris Huds.), tall fescue (Festuca arundinaceae Schreb.), or bare soil. ethofumesate [(+/-)-2-ethoxy-2,3-dihydro-3,3-dimethylbenzofuran-5-yl methansulfonate] was applied at 840 g ai ha(-)(1) on September 21, 1997. Halofenozide (N-4-chlorobenzoyl-N'-benzoyl-N'-tert-butylhydrazine) was applied at 1680 g ai ha(-)(1) on August 30, 1998. Replicate sampling cylinders were removed 2 h after treatment and 4, 8, 16, 32, and 64 days after treatment. Sampling cylinders were sectioned by depths and soil extracts were assayed by HPLC with a pesticide detection limit of 0.01 mg kg(-)(1). Turfgrass was divided into verdure and thatch and analyzed separately. ethofumesate leaching in turfgrass was reduced by at least 95% compared to leaching in bare soil. The half-life of ethofumesate in bare soil was 51 days compared to 3 days in turfgrass. Halofenozide showed similar leaching with or without turfgrass. Fifty percent dissipation of halofenozide did not occur within 64 days, regardless of organic matter cover.

Benzoates↗

Lignin deposition and associated changes in anatomy, enzyme activity, gene expression, and ruminal degradability in stems of tall fescue at different developmental stages.

Stem tissues of tall fescue (Festuca arundinacea Schreb.) were sampled at three elongation stages and three reproductive stages. Anatomical analysis showed the deposition of guaiacyl (G) and syringyl (S) lignin during plant development and the formation of a lignified sclerenchyma ring. A dramatic increase in Klason lignin content was found from elongation stage to reproductive stage. Lignin composition analyzed by gas chromatography-mass spectrometry revealed that S lignin content and S/G ratio increased with stem development, but contents of p-hydroxyphenyl (H) and G lignins decreased during the same period. S lignin content and S/G ratio also increased from the younger upper internode down to the older basal internode of the stem, but G and H lignin decreased in parallel. Relative O-methyltransferase activities increased during stem development and in parallel with the lignification process of stem. The pattern of enzyme activity during development varied with the choice of substrate, with highest activities seen when substrates were caffeoylaldehyde and 5-hydroxyferulic acid, and lowest activities were seen when caffeic acid and 5-hydroxyconiferyl alcohol were used as substrates. The expression of caffeic acid O-methyltransferase and cinnamyl alcohol dehydrogenase genes increased during the stem elongation stage and remained at high levels during the reproductive stages. The changes at anatomical, metabolic, and molecular levels during plant development were closely associated with lignification and degradability. This study provides an integrated picture of the molecular and chemical events that accompany changes in lignin deposition and ruminal degradability.

Alcohol Oxidoreductases↗

Contrasting responses of photosynthesis and carbon metabolism to low temperatures in tall fescue and clovers.

Growth, photosynthesis and carbohydrate metabolism in plants of two grassland species, clover (Trifolium subterraneum L. cv. Areces and Gaitan) and tall fescue (Festuca arundinacea Schreb.), shifted from 25 to 12 degrees C for 1 day or developed at 12 degrees C were compared with controls kept at 25 degrees C. Cold development produced a larger inhibition of growth in fescue than in clovers. In contrast, transferring plants from high to low temperature inhibited photosynthesis to a lesser extent in fescue than in clovers, this difference being associated with an increase in the activation state of Calvin cycle enzymes in fescue, but not in the clovers, a decreased cytosolic fructose-1,6-bisphosphatase (cFBPase, EC 3.1.3.11) activity in clovers, and an accumulation of hexose phosphates only in fescue. Development at 12 degrees C partly relieved the inhibition of photosynthesis in clovers, in contrast with fescue, which correlated with increases in total ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco, EC 4.1.1.39) activity only in clovers, and with greater increases in total stromal FBPase (sFBPase) activity in clovers than in fescue. The activity of sucrose synthesis enzymes was increased in the two clovers and fescue developed in the cold, while carbohydrate accumulation was much bigger in cold-developed fescue than in clovers because of a 5-fold increase in fructan contents in the former. The contents of phosphorylated intermediates increased in clovers but decreased in fescue grown at 12 degrees C. Our results suggest that restricted ribulose-1,5-bisphosphate (RuBP) regeneration limited the recovery of photosynthetic capacity in cold-developed fescue.

Journal Article↗