Introduction of the human lactoferrin gene into grass carp (Ctenopharyngodon idellus) to increase resistance against GCH virus

Haemorrhage can be an epidemic and fatal condition in grass carp. It is known now that the Grass Carp Haemorrhage Virus (GCHV) triggers haemorrhage. Human lactoferrin (hLF) plays an important role in the non-specific immune system, making some organisms more resistant to some viruses. Sperm of grass carp was mixed with linearized pCAhLFc, which is a DNA construct containing an hLF cDNA and the promoter of common carp beta-actin gene, and then electroporated. Then, mature eggs were fertilized in vitro with the treated sperm cells. The fry were sampled and analyzed by polymerase chain reaction (PCR). Results indicated that the foreign gene had been transferred successfully into the cells of some fry. Under optimal electroporation conditions, the efficiency of gene transfer was as high as 46.8%. About 35.7% of treated 5-month-old grass carp contained foreign genes. Most transgenic fry demonstrated significant delays in onset of symptoms of haemerrhage after injection of GCHV, suggesting a significant positive relationship between hLF cDNA and levels of disease resistance (P < 0.01). Results suggest that transgenic grass carp could be bred for increased resistance to haemorrhage. (C) 2002 Elsevier Science B.V. All rights reserved.

Effect of annual weeds on the growth of perennial grass mixtures in the alpine region of the Qinghai-Tibetan Plateau

In the alpine region of the Tibetan Plateau, five perennial grass cultivars, Bromus inermis (B), Elymus nutans (E), Clinelymus nutans (C), Agropyron cristatum (A), and Poa crymophila (P) were combined into nine communities with different compositions and ratios, B+C, E+A, B+E+A, E+B+C,C+E+A,B+E+C+A,B+C+A+P,B+E+A+P and E+C+A+P. Each combination was sown in six 10 X 10 m plots with three hand-weeded plots and three natural-growing plots in a completely randomised design in 1998. A field experiment studied the performance of these perennial grass combinations under the competitive interference of annual weeds in 3 consecutive years from 1998 to 2000. The results showed that annual weeds occupied more space and suppressed the growth of the grasses due to earlier germination and quicker growth in the establishment year, but this pattern changed in the second and third years. Leaf area indexes (LAIs) of grasses were greatly decreased by the competitive interference of weeds, and the negative effect of weeds on LAIs of grasses declined and stabilised in the second and third years. E+B+C, B+E+C+A, and B+E+A+P possessed relatively higher LAIs (P < 0.05) among all grass combinations and their LAIs were close to five when the competitive interference of weeds was removed. Grasses were competitively inferior to weeds in the establishment year, although their competitive ability (aggressivities) increased throughout the growing season. In the second and third years, grasses were competitively superior to weeds, and their competitive ability decreased from May until August and increased in September. Dry matter (DM) yields of grasses were reduced by 29.8-74.1% in the establishment year, 11.0-64.9% in the second year, and 16.0-55.8% in the third year by the competitive interference of weeds. B+E+C+A and B+E+A+P can produce around 14 t/ha of DM yields, significantly higher (P < 0.05) than the production of the other grass combinations in the second and third years after the competitive interference of weeds was removed. It was preliminarily concluded that removal of competitive interference of weeds increased the LAIs of all grass swards and improved the light interception of grasses, thus promoting the production of perennial grass pastures. The germination stage of the grasses in the establishment year was the critical period for weeding and suppression of weeds should occur at an early stage of plant growth. The grass combinations of B+E+C+A and B+E+A+P were productive and can be extensively established in the alpine regions of the Tibetan Plateau. Two or three growing seasons will be needed before determining success of establishment of grass mixtures under the alpine conditions of the Tibetan Plateau.

Productivity and persistence of perennial grass mixtures under competition from annual weeds in the alpine region of the Qinghai-Tibetan Plateau

In the alpine region of the Qinghai-Tibetan Plateau four indigenous perennial grass species Bromus inermis (BI), Elymus sibiricus (ES), Elymus nutans (EN) and Agropyron cristatum (AC) were cultivated as three mixtures with different compositions and seeding rates, BI + EN, BI + ES + AC and BI + ES + EN + AC. From 1998 to 2001 there were three different weeding treatments: never weeded (CK); weeded on three occasions in the first year (1-y) and weeded on three occasions in both the first and second year (2-y) and their effect of grass combination and interactions on sward productivity and persistence was measured. Intense competitive interference by weedy annuals reduced dry matter (DM) yield of the swards. Grass combination significantly affected sward DM yields, leaf area index (LAI) and foliar canopy cover and also species composition DM and LAI, and species plant cover. Interaction between weeding treatments and grass combination was significant for sward DM yield, LAI and canopy cover, but not on species composition for DM, LAI or species plant cover. Grass mixture BI + ES + EN + AC gave the highest sward DM yield and LAI for both weeding and non-weeding treatments. Species ES and EN were competitively superior to the others. Annual weedy forbs must be controlled to obtain productive and stable mixtures of perennial grasses, and germination/emergence is the most important time for removal. Weeding three times (late May, late June and mid-July) in the establishment year is enough to maintain the production and persistence of perennial grass mixtures in the following growing seasons. Extra weeding three times in the second growing year makes only a slight improvement in productivity.

The effect of feedstock cost on biofuel cost as exemplified by biomethane production from grass silage.

Abstract: The potential variance in feedstock costs can have signifi cant implications for the cost of a biofuel and the fi nancial viability of a biofuel facility. This paper employs the Grange Feed Costing Model to assess the cost of on-farm biomethane production using grass silages produced under a range of management scenarios. These costs were compared with the cost of wheat grain and sugarbeet roots for ethanol production at an industrial scale. Of the three feedstocks examined, grass silage represents the cheapest feedstock per GJ of biofuel produced. At a production cost of €27/tonne (t) feedstock (or €150/t volatile solids (VS)), the feedstock production cost of grass silage per gigajoule (GJ) of biofuel (€12.27) is lower than that of sugarbeet (€16.82) and wheat grain (€18.61). Grass biomethane is also the cheapest biofuel when grass silage is costed at the bottom quartile purchase price of silage of €19/t (€93/t VS). However, when considering the production costs (full-costing) of the three feedstocks, the total cost of grass biomethane (€32.37/GJ of biofuel; intensive 2-cut system) from a small on-farm facility ranks between that of sugarbeet (€29.62) and wheat grain ethanol (€34.31) produced in large industrial facilities. The feedstock costs for the above three biofuels represent 0.38, 0.57, and 0.54 of the total biofuel cost. The importance of feedstock cost on biofuel cost is further highlighted by the 0.43 increase in the cost of biomethane when grass silage is priced at the top quartile (€46/t or €232/t VS) compared to the bottom quartile purchase price.

Temporal and Georaphical Variations in Grass Pollen Seasons in Areas of Western Europe: an Analysis of Season dates at Sites of the European Pollen Information System.

Geographical and temporal variations in the start dates of grass pollen seasons are described for selected sites of the European Pollen Information Service. Daily average grass pollen counts are derived from Network sites in Finland, the Netherlands, Denmark, United Kingdom, Austria, Italy and Spain, giving a broad longitudinal transect over Western Europe. The study is part of a larger project that also examines annual and regional variations in the severity, timing of the peak and duration of the grass pollen seasons. For several sites, data are available for over twenty years enabling long term trends to be discerned. The analyses show notable contrasts in the progression of the seasons annually with differing lag times occurring between southern and northern sites in various years depending on the weather conditions. The patterns identified provide some insight into geographical differences and temporal trends in the incidence of pollinosis. The paper discusses the main difficulties involved in this type of analysis and notes possibilities for using data from the European Pollen Information service to construct pan European predictive models for pollen seasons.

Influence of the North Atlantic Oscillation on Grass Pollen Counts in Europe

Relationships between temporal variations in the North Atlantic Oscillation (NAO) and grass pollen counts at 13 sites in Europe, ranging from Córdoba in the South-West and Turku in the North-East, were studied in order to determine spatial differences in the amount of influence exerted by the NAO on the timing and magnitude of grass pollen seasons. There were a number of significant (p<0.05) relationships between the NAO and start dates of the grass pollen season at the 13 pollen-monitoring sites. The strongest associations were generally recorded near to the Atlantic coast. Several significant correlations also existed between winter averages of the NAO and grass pollen season severity. Traditional methods for predicting the start or magnitude of grass pollen seasons have centred on the use of local meteorological observations, but this study has shown the importance of considering large-scale patterns of climate variability like the NAO.

Digestion, rumen fermentation and circulating concentrations of insulin, growth hormone and IGF-1 in steers fed diets based on different proportions of maize silage and grass silage

Replacing grass silage with maize silage results in a fundamental change in the ratio of structural to non-structural carbohydrates with commensurate changes in rumen fermentation patterns and nutrient utilisation. This study investigated the effects of feeding four forage mixtures, namely grass silage (G); 67 g/100 g grass silage133 g/100 g maize silage (GGM); 67 g/100 g maize silage133/100 g grass silage (MMG); maize silage (M) to four ruminally and duodenally canulated Holstein Friesian steers. All diets were formulated to be isonitrogenous (22.4 g N/kg DM) using a concentrate mixture. Dietary dry matter (DM) and organic matter (OM) digestibility increased with ascending maize silage inclusion (P,0.1) whereas starch and neutral detergent fibre digestibility declined (P,0.05). Ratio of non-glucogenic to glucogenic precursors in the rumen fluid increased with maize silage inclusion (P,0.01) with a commensurate reduction in rumen pH (P,0.05). Mean circulating concentrations of insulin were greatest and similar in diets MMG and GGM, lower in diet M and lowest in diet G (P,0.01). There were no effects of diet on the mean circulating concentration of growth hormone (GH), or the frequency, amplitude and duration of GH pulses, or the mean circulating concentrations of IGF-1. Increasing levels of DM, OM and starch intakes with the substitution of grass silage with maize silage affected overall digestion, nutrient partitioning and subsequent circulating concentrations of insulin.

Apparent digestibility and nitrogen utilisation of diets based on maize and grass silage fed to beef steers

Substituting grass silage with maize silage in forage mixtures may result in one forage influencing the nutritive value of another in terms of whole tract nutrient digestibility and N utilisation. This experiment investigated effects of four forage combinations being, grass silage (G); 67 g/100 g grass silage + 33 g/100 g maize silage (GGM); 67 g/100 g maize silage + 33 g/100 g grass silage (MMG); maize silage (M). All diets were formulated to be isonitrogenous (22.4 g N/kg dry matter [DM]) using a concentrate mixture. Ration digestibility and N balance was determined using 7 Holstein Friesian steers (mean body weight 411.0 +/- 120.9 kg) in a cross-over design. Inclusion of maize silage in the diet had a positive linear effect on forage and total DM intake (P = 0.001), and on apparent DM and organic matter digestibility (both P = 0.048). Regardless of the silage ratio used, the metabolisable energy concentration of maize silage was calculated to be higher than that of grass silage (P = 0.058), and linearly related to the relative proportions of the two silages in the forage mixture. Inclusion of maize silage in the diet resulted in a linear decline in the apparent digestibility of starch (P = 0.022), neutral detergent fibre (P < 0.001) and acid detergent fibre (P = 0.003). Nitrogen retention, expressed as amount retained per day or in terms of body weight (g/100 kg) increased linearly with maize inclusion (P = 0.047 and 0.046, respectively). Replacing grass silage with maize silage caused linear responses according to the proportions of each forage in the diet, and that there were no associative effects of combining forages. (C) 2004 Elsevier B.V. All rights reserved.

Comparison of energy evaluation systems and a mechanistic model for milk production by dairy cattle offered fresh grass-based diets

Grass-based diets are of increasing social-economic importance in dairy cattle farming, but their low supply of glucogenic nutrients may limit the production of milk. Current evaluation systems that assess the energy supply and requirements are based on metabolisable energy (ME) or net energy (NE). These systems do not consider the characteristics of the energy delivering nutrients. In contrast, mechanistic models take into account the site of digestion, the type of nutrient absorbed and the type of nutrient required for production of milk constituents, and may therefore give a better prediction of supply and requirement of nutrients. The objective of the present study is to compare the ability of three energy evaluation systems, viz. the Dutch NE system, the agricultural and food research council (AFRC) ME system, and the feed into milk (FIM) ME system, and of a mechanistic model based on Dijkstra et al. [Simulation of digestion in cattle fed sugar cane: prediction of nutrient supply for milk production with locally available supplements. J. Agric. Sci., Cambridge 127, 247-60] and Mills et al. [A mechanistic model of whole-tract digestion and methanogenesis in the lactating dairy cow: model development, evaluation and application. J. Anim. Sci. 79, 1584-97] to predict the feed value of grass-based diets for milk production. The dataset for evaluation consists of 41 treatments of grass-based diets (at least 0.75 g ryegrass/g diet on DM basis). For each model, the predicted energy or nutrient supply, based on observed intake, was compared with predicted requirement based on observed performance. Assessment of the error of energy or nutrient supply relative to requirement is made by calculation of mean square prediction error (MSPE) and by concordance correlation coefficient (CCC). All energy evaluation systems predicted energy requirement to be lower (6-11%) than energy supply. The root MSPE (expressed as a proportion of the supply) was lowest for the mechanistic model (0.061), followed by the Dutch NE system (0.082), FIM ME system (0.097) and AFRCME system(0.118). For the energy evaluation systems, the error due to overall bias of prediction dominated the MSPE, whereas for the mechanistic model, proportionally 0.76 of MSPE was due to random variation. CCC analysis confirmed the higher accuracy and precision of the mechanistic model compared with energy evaluation systems. The error of prediction was positively related to grass protein content for the Dutch NE system, and was also positively related to grass DMI level for all models. In conclusion, current energy evaluation systems overestimate energy supply relative to energy requirement on grass-based diets for dairy cattle. The mechanistic model predicted glucogenic nutrients to limit performance of dairy cattle on grass-based diets, and proved to be more accurate and precise than the energy systems. The mechanistic model could be improved by allowing glucose maintenance and utilization requirements parameters to be variable. (C) 2007 Elsevier B.V. All rights reserved.

Comparison of heat-treated rapeseed expeller and solvent-extracted soya-bean meal as protein supplements for dairy cows given grass silage-based diets

Sixteen early to mid lactation Finnish Ayrshire dairy cows were used in a cyclic change-over experiment with four 21-day experimental periods and a 4 5 2 factorial arrangement of treatments to evaluate the effects of heat-treated rapeseed expeller and solvent-extracted soya-bean meal protein supplements on animal performance. Dietary treatments consisted of grass silage offered ad libitum supplemented with a fixed amount of a cereal based concentrate (10 kg/day on a fresh weight basis) containing 120, 150, 180 or 210 g crude protein (CP) per kg dry matter (DM). Concentrate CP content was manipulated by replacement of basal ingredients (g/kg) with either rapeseed expeller (R; 120, 240 and 360) or soya-bean meal (S; 80, 160 and 240). Increases in concentrate CP stimulated linear increases (P < 0.05) in silage intake (mean 22.5 and 23.8 g DM per g/kg increase in dietary CP content, for R and S, respectively) and milk production. Concentrate inclusion of rapeseed expeller elicited higher (P < 0.01) milk yield and milk protein output responses (mean 108 and 3.71 g/day per g/kg DM increase in dietary CP content) than soya-bean meal (corresponding values 62 and 2.57). Improvements in the apparent utilization of dietary nitrogen for milk protein synthesis (mean 0.282 and 0.274, for R and S, respectively) were associated with higher (P < 0.05) plasma concentrations of histidine, branched-chain, essential and total amino acids (35, 482, 902 and 2240 and 26, 410, 800 and 2119 mu mol/l, respectively) and lower (P < 0.01) concentrations of urea (corresponding values 4.11 and 4.52 mmol/l). Heat-treated rapeseed expeller proved to be a more effective protein supplement than solvent-extracted soya-bean meal for cows offered grass silage-based diets.

Simulating the effects of grassland management and grass ensiling on methane emission from lactating cows

A dynamic, mechanistic model of enteric fermentation was used to investigate the effect of type and quality of grass forage, dry matter intake (DMI) and proportion of concentrates in dietary dry matter (DM) on variation in methane (CH(4)) emission from enteric fermentation in dairy cows. The model represents substrate degradation and microbial fermentation processes in rumen and hindgut and, in particular, the effects of type of substrate fermented and of pH oil the production of individual volatile fatty acids and CH, as end-products of fermentation. Effects of type and quality of fresh and ensiled grass were evaluated by distinguishing two N fertilization rates of grassland and two stages of grass maturity. Simulation results indicated a strong impact of the amount and type of grass consumed oil CH(4) emission, with a maximum difference (across all forage types and all levels of DM 1) of 49 and 77% in g CH(4)/kg fat and protein corrected milk (FCM) for diets with a proportion of concentrates in dietary DM of 0.1 and 0.4, respectively (values ranging from 10.2 to 19.5 g CH(4)/kg FCM). The lowest emission was established for early Cut, high fertilized grass silage (GS) and high fertilized grass herbage (GH). The highest emission was found for late cut, low-fertilized GS. The N fertilization rate had the largest impact, followed by stage of grass maturity at harvesting and by the distinction between GH and GS. Emission expressed in g CH(4)/kg FCM declined oil average 14% with an increase of DMI from 14 to 18 kg/day for grass forage diets with a proportion of concentrates of 0.1, and on average 29% with an increase of DMI from 14 to 23 kg/day for diets with a proportion of concentrates of 0.4. Simulation results indicated that a high proportion of concentrates in dietary DM may lead to a further reduction of CH, emission per kg FCM mainly as a result of a higher DM I and milk yield, in comparison to low concentrate diets. Simulation results were evaluated against independent data obtained at three different laboratories in indirect calorimetry trials with COWS consuming GH mainly. The model predicted the average of observed values reasonably, but systematic deviations remained between individual laboratories and root mean squared prediction error was a proportion of 0.12 of the observed mean. Both observed and predicted emission expressed in g CH(4)/kg DM intake decreased upon an increase in dietary N:organic matter (OM) ratio. The model reproduced reasonably well the variation in measured CH, emission in cattle sheds oil Dutch dairy farms and indicated that oil average a fraction of 0.28 of the total emissions must have originated from manure under these circumstances.

Towards a lawn without grass: the journey of the imperfect lawn and its analogues

In much of the English-speaking world the lawn is the most common of all garden features. For arguably a millennium it has played a significant role in the landscape and during that period it has been inextricably linked with grasses. Nevertheless other plant species have accompanied the grasses and also been used in creating lawns. From medieval wildflowers to Victorian weeds, the plants that challenge the formal concept of the perfect lawn have journeyed with it but have until recently remained only small players within the dominion of grass. By the beginning of the 21st century, with a new environmental ethos permeating the garden, the long journey of the grassy lawn and its plant companions has led to the grass monoculture being heretically rethought: by removing both the monoculture and the grass.

The influence of plant species number on productivity, ground coverage and floral performance in grass-free lawns

The grass-free lawn is a novel development in modern ornamental horticulture where the traditional monoculture of grass is replaced by a variety of mowing-tolerant clonal forbs. It brings floral aesthetics and a diverse species approach to the use of lawn space. How the number of constituent forb species affects the aesthetic and structural performance of grass-free lawns was investigated using grass-free lawns composed of four, six and twelve British native clonal perennial forb species. Lawn productivity was seen to increase with increasing species number but the relationship was not linear. Plant cover was dynamic in all lawn types, varied between years and was not representative of individual species' floral performance. The behaviour of component species common to all lawns suggested that lawns with 12 species show greater structural stability than the lawns with a lower species number. Visual performance in lawns with the greatest species number was lower than in lawns with fewer species, with increasing variety in floral size and individual species floral productivity leading to a trade-off between diversity and floral performance. Individual species were seen to have different aesthetic functions in grass-free lawns either by providing flowers, ground coverage or both.

Prediction of metabolisable energy concentrations of fresh cut grass using digestibility data measured with non-pregnant non-lactating cows

Pasture-based ruminant production systems are common in certain areas of the world, but energy evaluation in grazing cattle is performed with equations developed, in their majority, with sheep or cattle fed total mixed rations. The aim of the current study was to develop predictions of metabolisable energy (ME) concentrations in fresh-cut grass offered to non-pregnant non-lactating cows at maintenance energy level, which may be more suitable for grazing cattle. Data were collected from three digestibility trials performed over consecutive grazing seasons. In order to cover a range of commercial conditions and data availability in pasture-based systems, thirty-eight equations for the prediction of energy concentrations and ratios were developed. An internal validation was performed for all equations and also for existing predictions of grass ME. Prediction error for ME using nutrient digestibility was lowest when gross energy (GE) or organic matter digestibilities were used as sole predictors, while the addition of grass nutrient contents reduced the difference between predicted and actual values, and explained more variation. Addition of N, GE and diethyl ether extract (EE) contents improved accuracy when digestible organic matter in DM was the primary predictor. When digestible energy was the primary explanatory variable, prediction error was relatively low, but addition of water-soluble carbohydrates, EE and acid-detergent fibre contents of grass decreased prediction error. Equations developed in the current study showed lower prediction errors when compared with those of existing equations, and may thus allow for an improved prediction of ME in practice, which is critical for the sustainability of pasture-based systems.

Prediction of nutrient digestibility and energy concentrations in fresh grass using nutrient composition

Improved nutrient utilization efficiency is strongly related to enhanced economic performance and reduced environmental footprint of dairy farms. Pasture-based systems are widely used for dairy production in certain areas of the world, but prediction equations of fresh grass nutritive value (nutrient digestibility and energy concentrations) are limited. Equations to predict digestible energy (DE) and metabolizable energy (ME) used for grazing cattle have been either developed with cattle fed conserved forage and concentrate diets or sheep fed previously frozen grass, and the majority of them require measurements less commonly available to producers, such as nutrient digestibility. The aim of the present study was therefore to develop prediction equations more suitable to grazing cattle for nutrient digestibility and energy concentrations, which are routinely available at farm level by using grass nutrient contents as predictors. A study with 33 nonpregnant, nonlactating cows fed solely fresh-cut grass at maintenance energy level for 50 wk was carried out over 3 consecutive grazing seasons. Freshly harvested grass of 3 cuts (primary growth and first and second regrowth), 9 fertilizer input levels, and contrasting stage of maturity (3 to 9 wk after harvest) was used, thus ensuring a wide representation of nutritional quality. As a result, a large variation existed in digestibility of dry matter (0.642-0.900) and digestible organic matter in dry matter (0.636-0.851) and in concentrations of DE (11.8-16.7 MJ/kg of dry matter) and ME (9.0-14.1 MJ/kg of dry matter). Nutrient digestibilities and DE and ME concentrations were negatively related to grass neutral detergent fiber (NDF) and acid detergent fiber (ADF) contents but positively related to nitrogen (N), gross energy, and ether extract (EE) contents. For each predicted variable (nutrient digestibilities or energy concentrations), different combinations of predictors (grass chemical composition) were found to be significant and increase the explained variation. For example, relatively higher R(2) values were found for prediction of N digestibility using N and EE as predictors; gross-energy digestibility using EE, NDF, ADF, and ash; NDF, ADF, and organic matter digestibilities using N, water-soluble carbohydrates, EE, and NDF; digestible organic matter in dry matter using water-soluble carbohydrates, EE, NDF, and ADF; DE concentration using gross energy, EE, NDF, ADF, and ash; and ME concentration using N, EE, ADF, and ash. Equations presented may allow a relatively quick and easy prediction of grass quality and, hence, better grazing utilization on commercial and research farms, where nutrient composition falls within the range assessed in the current study.