Higher Water-soluble Carbohydrate Research Articles

Natural lactic acid bacteria population of tropical grasses and their fermentation factor analysis of silage prepared with cellulase and inoculant.

Natural lactic acid bacteria (LAB) populations in tropical grasses and their fermentation characteristics on silage prepared with cellulase enzyme and LAB inoculants were studied. A commercial inoculant Lactobacillus plantarum Chikuso 1 (CH), a local selected strain Lactobacillus casei TH14 (TH14), and 2 cellulases, Acremonium cellulase (AC) and Maicelase (MC; Meiji Seika Pharma Co. Ltd., Tokyo, Japan), were used as additives to silage preparation with fresh and wilted (6 h) Guinea grass and Napier grass. Silage was prepared using a laboratory-scale fermentation system. Treatments were CH, TH14, AC at 0.01% fresh matter, AC 0.1%, MC 0.01%, MC 0.1%, CH+AC 0.01%, CH+AC 0.1%, CH+MC 0.01%, CH+MC 0.1%, TH14+AC 0.1%, TH14+AC 0.01%, TH14+MC 0.1%, and TH14+MC 0.01%. Microorganism counts of Guinea grass and Napier grass before ensiling were 102 LAB and 106 aerobic bacteria; these increased during wilting. Based on morphological and biochemical characteristics, and 16S rRNA gene sequence analysis, natural strains from both grasses were identified as L. plantarum, L. casei, Lactobacillus acidipiscis, Leuconostoc pseudomesenteroides, Leuconostoc garlicum, Weissella confusa, and Lactococcus lactis. Lactobacillus plantarum and L. casei are the dominant species and could grow at lower pH and produce more lactic acid than the other isolates. Crude protein and neutral detergent fiber were 5.8 and 83.7% of dry matter (DM) for Guinea grass, and 7.5 and 77.1% of DM for Napier grass. Guinea grass had a low level of water-soluble carbohydrates (0.39% of DM). Guinea grass silage treated with cellulase had a lower pH and higher lactic acid content than control and LAB treatments. The 0.1% AC and MC treatments had the best result for fermentation quality. All high water-soluble carbohydrate (2.38% DM) Napier grass silages showed good fermentation quality. Compared with control and LAB-inoculated silage, the cellulase-treated silages had significantly higher crude protein content and lower neutral detergent fiber and acid detergent fiber contents. The results confirmed that cellulase could improve tropical silage quality, inhibiting protein degradation and promoting fiber degradation.

Genotypic variation in the accumulation of water-soluble carbohydrate in canola and its potential contribution to seed yield in different environments

Water-soluble carbohydrates (WSC) stored in the stems and pod wall in canola (Brassica napus L.) can be an important source of assimilates contributing to yield. However, genotypic variation in WSC and its contribution to yield have not been investigated. The aims of this work were to quantify genotypic variation of WSC and the extent to which WSC contributes to seed yield. We measured the concentration and content of WSC in a number of hybrid and open pollinated triazine tolerant (TT) and non-TT canola genotypes at flowering, podding and maturity from 2009 to 2011. There were significant repeatable differences in WSC concentration among genotypes (28–108mgg−1 at podding) and WSC content (12–113gm−2 at podding). The major difference in WSC content occurred between the TT and non-TT canola while the difference between hybrid and open-pollinated canola was small. On average, non-TT canola produced higher amount of WSC (68gm−2) than TT canola (47gm−2). Seed yield increased with the increased amount of remobilized WSC in the limited water-stressed conditions (2009 and 2011). Despite the high amount of WSC in drought (2010) year, no significant relationship was found between yield and the remobilized WSC because the higher WSC was offset by yield penalty from delay in flowering in these varieties. The contribution of WSC to yield was higher in drought (10–22%, 2010) year than in the average (3–9%, 2009) and above-average (7–12%, 2011) rainfall year. Genotype and environment interaction was relatively small compared to genotype effect. The difference in the amount of WSC between the herbicide tolerant groups and among genotypes was repeatable and the rankings of genotypes for WSC remained similar across the years. Both the WSC content and the ratio of remobilized WSC to yield had large broad-sense heritability (H=0.68–0.85). The high heritability, the positive relationships between seed yield and WSC under limited water stress conditions, and the high contribution of WSC to yield under drought conditions suggest that WSC could be used as a breeding trait to augment yield in the limited water-stress condition and maintain yield under drought conditions by reducing the effect of water stress on yield.

Effect of harvest date on Arundo donax L. (giant reed) composition, ensilage performance, and enzymatic digestibility

Composition and ensilage performance of giant reed harvested in August, October, November, and December, were evaluated and compared. Generally, late-harvested giant reed had higher dry matter content, lower nitrogen content, and higher water soluble carbohydrates (WSC) content than early-harvested giant reed. During 90days of ensilage, giant reed harvested in October, November, and December showed dry matter losses of about 1%, while giant reed harvested in August showed a higher dry matter loss of about 8%. During the ensilage process, more lactic acid was produced in late-harvested giant reed than in early-harvested giant reed. Late-harvested giant reed had a higher lignin content and lower enzymatic digestibility than early-harvested giant reed. However, enzymatic digestibility of all the giant reed biomass was improved by the 90-day ensilage process, reaching levels of 43–46%. In summary, ensilage could be used for storing giant reed biomass harvested at different times and for improving its digestibility.

Cultivar by environment effects of perennial ryegrass cultivars selected for high water soluble carbohydrates managed under differing precipitation levels

Historic results of perennial ryegrass (Lolium perenne L.) breeding include improved disease resistance, biomass, and nutritional quality. Yet, lack of tolerance to water stress limits its wide use. Recent efforts to increase water soluble carbohydrate (WSC) content in perennial ryegrass may increase drought tolerance. Herein, we report results of a multi-year and location evaluation of differing precipitation/irrigations levels on genetic and genotype × environment interaction effects of various agronomic traits in perennial ryegrass. The study included two UK environments (Aberystwyth, Wales and Edinburgh, Scotland) and four USA environments located in Logan, UT and consisting in four supplemental irrigation levels in a line-source irrigation design. Data included herbage dry matter, dry matter digestibility, crude protein, and WSC, collected from 2011 to 2013. There were differences (P < 0.05) among the included cultivars for each phenotype when evaluated across environments. There was observed cultivar × environment interaction for each phenotype, which precluded the identification of the best cultivar over all environments. Additive main effect and multiplicative interaction modeling allowed for grouping of the individual environments to mega-environments. The analysis identified three mega-environments for each phenotype: (1) the UK environments, (2) the two highest Logan irrigation levels, and (3) the two lowest Logan irrigation levels. Mega-environments designations corresponded to water stress, temperature, and inherent geographic/climate differences between the environments. The evidence for the potential of increased WSC to improve drought tolerance under water stress was inconclusive, but high WSC cultivars also exhibited high herbage production under the imposed water stress.

Fructan synthesis, accumulation and polymer traits. II. Fructan pools in populations of perennial ryegrass (Lolium perenne L.) with variation for water-soluble carbohydrate and candidate genes were not correlated with biosynthetic activity and demonstrated constraints to polymer chain extension.

Differences have been shown between ryegrass and fescue within the Festulolium subline introgression family for fructan synthesis, metabolism, and polymer-size traits. It is well-established that there is considerable variation for water-soluble carbohydrate and fructan content within perennial ryegrass. However there is much still to be discovered about the fructan polymer pool in this species, especially in regard to its composition and regulation. It is postulated that similar considerable variation for polymer traits may exist, providing useful polymers for biorefining applications. Seasonal effects on fructan content together with fructan synthesis and polymer-size traits have been examined in diverse perennial ryegrass material comprising contrasting plants from a perennial ryegrass F2 mapping family and from populations produced by three rounds of phenotypic selection. Relationships with copy number variation in candidate genes have been investigated. There was little evidence of any variation in fructan metabolism across this diverse germplasm under these conditions that resulted in substantial differences in the complement of fructan polymers present in leaf tissue at high water-soluble carbohydrate concentrations. The importance of fructan synthesis during fructan accumulation was unclear as fructan content and polymer characteristics in intact plants during the growing season did not reflect the capacity for de novo synthesis. However, the retention of fructan in environmental conditions favoring high sink/low source demand may be an important component of the high sugar trait and the roles of breakdown and turnover are discussed.

Effect of water-soluble carbohydrate content in orchardgrass pasture on grazing time and rumen fermentation in dairy cows.

Two experiments were conducted to clarify the effect of water-soluble carbohydrate (WSC) content in orchardgrass pasture on the diurnal distribution of grazing time. Six ruminally cannulated, non-lactating dairy cows were grazed on either of two pastures with different orchardgrass cultivars containing low WSC (LWSC; cultivar: 'Hokkai 28') or high WSC (HWSC; cultivar: 'Harunemidori'). The cows were grazed in morning and evening sessions in experiment 1, whereas the cows were grazed throughout the day in experiment 2. In experiment 1, grazing time of the cows on HWSC was longer than that of the cows on LWSC (P < 0.01). This difference was larger in the morning session than in the evening session (pasture × grazing session: P < 0.05). Effects on herbage intake were similar to those on grazing time. In experiment 2, daily total grazing time was longer for the cows on HWSC than for those on LWSC (P < 0.05). The cows on HWSC spent a longer time grazing than those on LWSC in the morning between 03.00 and 09.00 hours (P < 0.01). The results indicated that prolonged grazing time in the period between dawn and early morning could increase daily herbage intake in cows grazed on pastures of orchardgrass cultivars with high-WSC content.

Dynamics Associated with Prolonged Ensiling and Aerobic Deterioration of Total Mixed Ration Silage Containing Whole Crop Corn.

This study investigated the dynamics associated with prolonged ensiling and aerobic deterioration of whole crop corn (WCC) silages and total mixed ration (TMR) silages containing WCC (C-TMR silages) to clarify the differences that account for the enhanced aerobic stability of TMR silages. Laboratory-scale barrel silos were randomly opened after 7, 14, 28, and 56 d of ensiling and were subjected to analyses of fermentation quality, microbial and temperature dynamics during aerobic exposure. WCC and C-TMR silages were both well preserved and microorganisms were inhibited with prolonged ensiling, including lactic acid bacteria. Yeast were inhibited to below the detection limit of 500 cfu/g fresh matter within 28 d of ensiling. Aerobic stability of both silages was enhanced with prolonged ensiling, whereas C-TMR silages were more aerobically stable than WCC silages for the same ensiling period. Besides the high moisture content, the weak aerobic stability of WCC silage is likely attributable to the higher lactic acid content and yeast count, which result from the high water-soluble carbohydrates content in WCC. After silo opening, yeast were the first to propagate and the increase in yeast levels is greater than that of other microorganisms in silages before deterioration. Besides, increased levels of aerobic bacteria were also detected before heating of WCC silages. The temperature dynamics also indicated that yeast are closely associated with the onset of the aerobic deterioration of C-TMR silage, whereas for WCC silages, besides yeast, aerobic bacteria also function in the aerobic deterioration. Therefore, the inclusion of WCC might contribute to the survival of yeast during ensiling but not influence the role of yeast in deterioration of C-TMR silages.

Favorable alleles for stem water-soluble carbohydrates identified by association analysis contribute to grain weight under drought stress conditions in wheat.

Drought is a major environmental constraint to crop distribution and productivity. Stem water-soluble carbohydrates (WSC) buffer wheat grain yield against conditions unfavorable for photosynthesis during the grain filling stage. In this study, 262 winter wheat accessions and 209 genome-wide SSR markers were collected and used to undertake association analysis based on a mixed linear model (MLM). The WSC in different internodes at three growth stages and 1000-grain weight (TGW) were investigated under four environmental regimes (well-watered, drought stress during the whole growth period, and two levels of terminal drought stress imposed by chemical desiccation under the well-watered and drought stress during the whole growth period conditions). Under diverse drought stress conditions, WSC in lower internodes showed significant positive correlations with TGW, especially at the flowering stage under well-watered conditions and at grain filling under drought stress. Sixteen novel WSC-favorable alleles were identified, and five of them contributed to significantly higher TGW. In addition, pyramiding WSC favorable alleles was not only effective for obtaining accessions with higher WSC, but also for enhancing TGW under different water regimes. During the past fifty years of wheat breeding, WSC was selected incidentally. The average number of favorable WSC alleles increased from 1.13 in the pre-1960 period to 4.41 in the post-2000 period. The results indicate a high potential for using marker-assisted selection to pyramid WSC favorable alleles in improving WSC and TGW in wheat.

Longer defoliation interval ensures expression of the ‘high sugar’ trait in perennial ryegrass cultivars in cool temperate Tasmania, Australia

SUMMARYPerennial ryegrass (Lolium perenneL.) cultivars have been developed to express higher levels of leaf water-soluble carbohydrates (WSC), but expression of this ‘high sugar’ trait varies between environments and is likely to be further influenced by the extent of plant re-growth. The herbage WSC concentration and the ratio of WSC to crude protein (WSC : CP) in high sugar cultivars AberMagic and SF Joule were therefore compared with a control cultivar, Arrow, under cool temperate Tasmanian conditions and two defoliation interval treatments. The irrigated cultivars were subjected to defoliation at either the 1·5-leaf or 3-leaf stage of re-growth, and additional components of nutritive value (fibre concentrations and metabolizable energy content) and dry matter (DM) yields were measured throughout a 12-month period (March 2011 to March 2012). The high sugar trait was consistently expressed in AberMagic, which under both the 1·5-leaf and 3-leaf stages defoliation intervals, displayed the highest WSC concentration (mean 194 and 247 mg/g DM, respectively, compared with 153 and 178 mg/g DM for Arrow) and highest WSC : CP ratio (mean 0·74 and 1·29, respectively, compared with 0·58 and 0·85 for Arrow). Defoliation at the 3-leaf stage of regrowth ensured greater expression of the high sugar trait in both AberMagic and SF Joule, as measured by the increase in WSC concentration and WSC : CP ratio as a result of increasing defoliation interval. The strength and consistency of trait expression in AberMagic under the 3-leaf stage defoliation interval warrants further research to investigate its effect on rumen nitrogen (N) partitioning and milk production in this environment.

Genetic Differentiation in Response to Selection for Water-Soluble Carbohydrate Content in Perennial Ryegrass (Lolium perenne L.)

Plant carbohydrates are of increasing interest as renewable feedstocks to replace petrochemicals in the generation of fuels and production of high-value chemicals. Greater understanding of the genetic control of diversity in fructan synthesis and accumulation would facilitate more directed channelling of feedstock to process in a ryegrass biorefinery. Divergent populations produced by phenotypic selection for water-soluble carbohydrate content have been used to investigate relationships between traits, and to identify patterns of genetic differentiation which indicate genomic regions under high and low selection pressure. Selection for high water-soluble carbohydrate content was associated with increased synthesis of large fructan polymers and increased accumulation of above-ground plant biomass, particularly during spring. Three rounds of selection and two rounds of recombination resulted in widespread genetic differentiation across the whole genome, causing reduced allelic richness and increasing homozygosity at some loci. A smaller number of loci were shown to be subject to high selection pressure. Breeding material subjected to many years of selection for water-soluble carbohydrate also showed allelic differences which may reflect the consequences of high selection pressure at some of these same loci. However, some of the loci unaffected in the divergent selection experiment showed similar effects. This might arise from differences in linkage disequilibrium in these two sets of plant materials, but more likely from the different genetic background of the germplasm. This illustrates the complex nature of the water-soluble carbohydrate trait in perennial ryegrass.

Efficient extraction of fructans from sotol plant (Dasylirion leiophyllum) enhanced by a combination of enzymatic and sonothermal treatments

The effects of different extraction methods of water-soluble carbohydrates (WSC) from the sotol plant (Dasylirion leiophyllum) were investigated. Sotol fragments were extracted at 40 and 70°C, under thermal treatment (T), pre-enzymatic thermal treatment (PET), sonothermal treatment (ST), and pre-enzymatic sonothermal treatment (PEST) conditions: fructose, glucose, sucrose, and fructans were analyzed by HPLC and the total water soluble carbohydrates was determined. At 70°C, the highest WSC values (482mg/gd.m.) were obtained, with a fructan proportion of 69%. Pre-enzymatic treatment at 70°C resulted in a high WSC content with the highest fructans proportion (87%) and lowest contents of RS and sucrose. The effect of the interaction between ultrasound and enzymatic treatments was limited by the high-temperature effect (70°C), thereby minimizing the extraction. Microscopy analyses showed cell-wall modifications with the ST and PET treatments, which caused an increase in the total soluble sugars. The combination of enzymatic and sonothermal treatments at 70°C resulted in the extraction of fructans in a higher yield and with less degradation. This circumvents the need for traditional high-energy processes, which could be beneficial for the extraction of WSC such as fructans from sotol or other economically important plants.

The potential of diverse pastures to reduce nitrogen leaching on New Zealand dairy farms

The largest contributor to nitrogen (N) leaching from ryegrass-clover pasture based dairy farms is the surplus feed N excreted as urinary N (UN) onto pastures. Pastures consisting of mixtures of ryegrass, herbs and legumes (diverse pastures) have shown potential to yield similar DM, but with a lower N content and a higher water soluble carbohydrate : crude protein ratio compared with standard ryegrass–clover pastures. These diverse pastures have shown the potential to lower the UN excreted by dairy cows in short-term, late-lactation studies. This modelling study was designed to scale the results from component studies up to farm and over a full season to evaluate the potential of diverse pastures to become a suitable strategy for reducing N leaching on New Zealand dairy farms. The Molly cow model was tested against observed data from one indoor and one outdoor study where feeding diverse pasture resulted in UN (N excreted in urine g/day) reductions of 50% and 17%, respectively. The model predicted UN reductions of 23% and 17%. Farm-scale model scenarios, where 20% or 50% of the farm was sown with diverse pastures, resulted in 2% and 6% reductions in UN deposited onto paddocks. This reduction was smaller than expected with some system interactions related to seasonal feed supply, diet composition and total N intake being likely to play a role. The reduction in UN onto paddocks, together with a dilution effect from larger urine volumes per cow per day as a result of lower DM% of diverse pastures, resulted in N leaching reductions of 11% and 19% for the two scenarios, respectively. This potential to reduce N leaching needs to be evaluated further in the context of farm profitability when other aspects of diverse pastures such as yield, persistency, drought resistance and ability to extract N from the soil becomes part of the farm-system analysis.

More fertile florets and grains per spike can be achieved at higher temperature in wheat lines with high spike biomass and sugar content at booting.

An understanding of processes regulating wheat floret and grain number at higher temperatures is required to better exploit genetic variation. In this study we tested the hypothesis that at higher temperatures, a reduction in floret fertility is associated with a decrease in soluble sugars and this response is exacerbated in genotypes low in water soluble carbohydrates (WSC). Four recombinant inbred lines contrasting for stem WSC were grown at 20/10°C and 11h photoperiod until terminal spikelet, and then continued in a factorial combination of 20/10°C or 28/14°C with 11h or 16h photoperiod until anthesis. Across environments, High WSC lines had more grains per spike associated with more florets per spike. The number of fertile florets was associated with spike biomass at booting and, by extension, with glucose amount, both higher in High WSC lines. At booting, High WSC lines had higher fixed 13C and higher levels of expression of genes involved in photosynthesis and sucrose transport and lower in sucrose degradation compared with Low WSC lines. At higher temperature, the intrinsic rate of floret development rate before booting was slower in High WSC lines. Grain set declined with the intrinsic rate of floret development before booting, with an advantage for High WSC lines at 28/14°C and 16h. Genotypic and environmental action on floret fertility and grain set was summarised in a model.

Relationship between Water Soluble Carbohydrate Content, Aphid Endosymbionts and Clonal Performance of Sitobion avenae on Cocksfoot Cultivars

Aphids feed on plant phloem sap, rich in sugars but poor in essential amino acids. However, sugars cause osmotic regulation problems for aphids, which they overcome by hydrolysing the sugars in their gut and polymerising the hydrolysis products into oligosaccharides, excreted with honeydew. Aphids harbour primary bacterial endosymbionts, which supply them with essential amino acids necessary for survival. They also harbour secondary (facultative) endosymbionts (sfS), some of which have a positive impact on life history traits, although it is not yet known whether they also play a role in providing effective tolerance to differing levels of water soluble carbohydrates (WSCs). We investigated the relationship between WSC content of cocksfoot cultivars and performance of clones of the English grain aphid Sitobion avenae F. We evaluated how clone genotype and their sfS modulate performance on these different cultivars. We therefore examined the performance of genetically defined clones of S. avenae, collected from different host plants, harbouring different sfS. The performance was tested on 10 Dactylis glomerata L. cultivars with varying WSC content. D. glomerata is known as a wild host plant for S. avenae and is also commercially planted. We found that high WSCs levels are responsible for the resistance of D. glomerata cultivars to specific S. avenae clones. The minimum level of WSCs conferring resistance to D. glomerata cultivars was 1.7% dw. Cultivars with a WSC content of 2.2% or higher were resistant to S. avenae and did not allow reproduction. Our results further indicate that sfS modulate to some extend host plant cultivar adaptation in S. avenae. This is the first study revealing the importance of WSCs for aphid performance. Cocksfoot cultivars with a high content of WSCs might be therefore considered for aphid control or used for resistance breeding in this and other grass species, including cereals.

Developmental and growth controls of tillering and water-soluble carbohydrate accumulation in contrasting wheat (Triticum aestivum L.) genotypes: can we dissect them?

In wheat, tillering and water-soluble carbohydrates (WSCs) in the stem are potential traits for adaptation to different environments and are of interest as targets for selective breeding. This study investigated the observation that a high stem WSC concentration (WSCc) is often related to low tillering. The proposition tested was that stem WSC accumulation is plant density dependent and could be an emergent property of tillering, whether driven by genotype or by environment. A small subset of recombinant inbred lines (RILs) contrasting for tillering was grown at different plant densities or on different sowing dates in multiple field experiments. Both tillering and WSCc were highly influenced by the environment, with a smaller, distinct genotypic component; the genotype×environment range covered 350–750 stems m–2 and 25–210mg g–1 WSCc. Stem WSCc was inversely related to stem number m–2, but genotypic rankings for stem WSCc persisted when RILs were compared at similar stem density. Low tillering–high WSCc RILs had similar leaf area index, larger individual leaves, and stems with larger internode cross-section and wall area when compared with high tillering–low WSCc RILs. The maximum number of stems per plant was positively associated with growth and relative growth rate per plant, tillering rate and duration, and also, in some treatments, with leaf appearance rate and final leaf number. A common threshold of the red:far red ratio (0.39–0.44; standard error of the difference=0.055) coincided with the maximum stem number per plant across genotypes and plant densities, and could be effectively used in crop simulation modelling as a ‘cut-off’ rule for tillering. The relationship between tillering, WSCc, and their component traits, as well as the possible implications for crop simulation and breeding, is discussed.

Yield and Quality Response of Perennial Ryegrass Selected for High Concentration of Water‐Soluble Carbohydrate to Nitrogen Application Rate

ABSTRACTThe expression of elevated water‐soluble carbohydrate (WSC) concentrations in perennial ryegrass (Lolium perenne L.) cultivars selected for high forage WSC concentration can be highly variable across environments. Our aim was to determine whether N application rate influences the expression of the high WSC phenotype. Cultivars AberDart (selected for high WSC concentration) and Fennema (control) were evaluated across four fertilizer N application rates (0, 40, 80, and 120 kg ha−1 per harvest) over four replicates and 2 yr at Grange, Ireland, and Særheim, Norway. Plots were managed for silage production with four cuts per year in Ireland and three cuts per year in Norway. Nine forage traits were measured: WSC, dry matter digestibility, crude protein, buffering capacity, dry matter, ash, dry matter yield, N use efficiency, and apparent N recovery. The response of AberDart and Fennema to N application rate was predominantly similar within and over years and locations for all traits. Differences between cultivars in WSC concentration were largely consistent across N application rates, years, and locations. AberDart had mean WSC concentrations 8 to 12% higher than Fennema depending on harvest. Present results suggest that the evaluation and selection of perennial ryegrass for high concentrations of WSC in cool‐temperate maritime climates may be conducted across a wide range of N application rates and, by extension, soil N supply rates.

Experimental methods for laboratory-scale ensilage of lignocellulosic biomass

Anaerobic fermentation is a potential storage method for lignocellulosic biomass in biofuel production processes. Since biomass is seasonally harvested, stocks are often dried or frozen at laboratory scale prior to fermentation experiments. Such treatments prior to fermentation studies cause irreversible changes in the plant cells, influencing the initial state of biomass and thereby the progression of the fermentation processes itself. This study investigated the effects of drying, refrigeration, and freezing relative to freshly harvested corn stover in lab-scale ensilage studies. Particle sizes, as well as post-ensilage drying temperatures for compositional analysis, were tested to identify the appropriate sample processing methods. After 21 days of ensilage the lowest pH value (3.73 ± 0.03), lowest dry matter loss (4.28 ± 0.26 g. 100 g-1DM), and highest water soluble carbohydrate (WSC) concentrations (7.73 ± 0.26 g. 100 g-1DM) were observed in control biomass (stover ensiled within 12 h of harvest without any treatments). WSC concentration was significantly reduced in samples refrigerated for 7 days prior to ensilage (3.86 ± 0.49 g. 100 g−1 DM). However, biomass frozen prior to ensilage produced statistically similar results to the fresh biomass control, especially in treatments with cell wall degrading enzymes. Grinding to decrease particle size reduced the variance amongst replicates for pH values of individual reactors to a minor extent. Drying biomass prior to extraction of WSCs resulted in degradation of the carbohydrates and a reduced estimate of their concentrations. The methods developed in this study can be used to improve ensilage experiments and thereby help in developing ensilage as a storage method for biofuel production.

Effect of feeding dried high-sugar ryegrass (‘AberMagic’) on methane and urinary nitrogen emissions of primiparous cows

The objective of the present study was to test the hypothesis that feeding dried grass from a ryegrass cultivar with high water-soluble carbohydrate (WSC) concentration to dairy cows lowers emissions of enteric methane and urinary nitrogen. This dried grass (here the cultivar ‘AberMagic’) was compared as a forage-only diet to dried grass prepared from a low-WSC cultivar (‘Respect’). Six mid-lactating primiparous Holstein-Friesian dairy cows (19kg milk/cow/day) were subjected to a change-over design in two 26-day experimental periods. In the last 8 days of each period, feed intake, milk yield, fecal and urinary excretion were recorded and samples were taken. During 2 days, methane emissions were measured in open-circuit respiration chambers. The dried grass type had no significant effect on feed intake and milk gross composition. Milk yield was lower for cows fed the dried high-WSC grass. Total enteric methane (g/day) did not significantly differ when cows received dried high-WSC grass (279) instead of dried low-WSC grass (315). Methane expressed as g/kg dry matter intake (19.4 and 20.3 for dried high- and low-WSC grass, respectively), and methane conversion rates (Ym; 63kJ/MJ of gross energy for both dried grass types) were similar with both dried grass types. The dried high-WSC grass had a lower crude protein concentration compared to the dried low-WSC grass (158 vs. 255g/kg dry matter) resulting in a correspondingly different urinary N excretion (114 vs. 292g/day) and milk urea concentration. A similar effect was found for urinary N proportion of total excreta N. Fecal N excretion and milk N secretion were higher with the dried high-WSC grass. In conclusion, feeding dried high-WSC ryegrass, where WSC mainly replace crude protein, might be helpful in limiting N emissions from dairy cattle husbandry whereas methane emissions seem to remain unaffected. Further studies have to test whether dried high-WSC grass has the same effect when the WSC replace fiber and to address the extent to which favorable effects can be recovered in mixed forage-concentrate diets.

Yield and chemical composition of five common grassland species in response to nitrogen fertiliser application and phenological growth stage

The interaction of grassland management factors such as plant species, rate of nitrogen (N) fertiliser application and stage of maturity at harvest, will determine the optimal balance of herbage yield, nutritive quality and ensilability for ruminant nutrition and/or industrial applications. This study investigated the effects of N fertiliser input and harvest date on the yield and chemical composition of five common grass species, and made comparisons with red clover. Perennial ryegrass (PRG), Italian ryegrass (IRG), tall fescue, cocksfoot, timothy and red clover were grown under two inorganic N fertiliser inputs (0 kg N ha−1 and 125 kg N ha−1; except red clover) and harvested at five dates (fortnightly from 12 May to 7 July) in the primary growth. Regression analysis of these data allowed comparison of the yield and chemical composition of each grass species at common growth stages, without the confounding effects of variation in maturity between grass species at common harvest dates. Of the grass species investigated, timothy was most productive in terms of dry matter (DM) yield and thus has the potential to provide a cheaper feed per unit DM. However, the most digestible grass species was PRG, with timothy being the lowest, and this could impact on both animal and bioenergy production potential. The most suitable grass species for ensiling was IRG (particularly when grown without fertiliser N) due to its higher water soluble carbohydrate (WSC) concentration and lower buffering capacity (BC) compared to all other grass species. In comparison to the grasses receiving inorganic N fertiliser, red clover had a numerically lower DM yield, but a higher mean DM digestibility and crude protein concentration. The lower WSC concentration and higher BC of the red clover may result in a greater preservation challenge during ensiling.

Effects of water soluble carbohydrate content on ensiling characteristics, chemical composition and in vitro gas production of forage millet and forage sorghum silages

A study was conducted to determine effects of water soluble carbohydrate (WSC) concentration on ensiling characteristics, chemical composition and in vitro digestibility of forage millet and forage sorghum. Two cultivars (regular and high WSC content) of each forage were harvested at late vegetative stage and ensiled in mini-silos for 0, 2, 4, 8, 16 and 48d. Ensiled forages went through a rapid fermentation as indicated by a sharp decline in pH during the first 2d of ensiling. Between day 0 and 8 post-ensiling, pH was higher (P<0.05) for ensiled millet than ensiled sorghum forages and was lower for high WSC than regular cultivars. Lactic acid concentration was higher (P<0.05) in high WSC than in regular cultivars and was not affected by forage type. Chemical composition of the 45-d silages showed that forage millet silages contain more (P<0.05) neutral (aNDF) and acid detergent fiber than forage sorghum silages. Fiber fractions were not affected by WSC content except for lignin(sa), which was higher (P<0.05) for regular versus high WSC cultivars. Soluble and non-protein N were higher (P<0.05) in forage millet than forage sorghum silages and were higher for high WSC versus normal cultivars. In vitro gas production, degradation rate and volatile fatty acid production were not affected by forage type but were higher (P<0.05) for high WSC versus normal cultivars. In vitro dry matter and aNDF digestibilities were higher for forage sorghum versus forage millet silages, and were higher for high WSC versus normal cultivars. High WSC content improved ensiling characteristics of both forages, but the effect was more pronounced for forage sorghum than for forage millet. Based on chemical composition and in vitro data, we conclude that the feeding value of forage sorghum silage is superior to that of forage millet silage when harvested at the same stage of physiological development.