Opportunities in Molecular Biology: Enhancement of the Nutritional Value of Forages

Pasture plants, while producing a cost-effective source of feed for grazing ruminants, can frequently be less than optimal in meeting the animals’ nutrient requirements. Over the past half century, there has been a major effort to improve the quantity and nutritional quality of pasture plants using conventional plant breeding. Although considerable progress has been made in improving the quality and agronomic characteristics of our major pasture plants by this means, breeding can only be applied to plants capable of sexual crosses. This poses severe limitations both in terms of speed of progress and in the number of genes available for transfer (Ulyatt, 1991; Ulyatt et al., 1995)

Rumen function and digestion parameters associated with differences between sheep in methane emissions when fed chaffed lucerne hay

An indoor experiment involving 10 rumen-cannulated Romney sheep was conducted in May and June 1998 at AgResearch Grasslands, Palmerston North, New Zealand, under restricted feeding conditions. in order to test the hypothesis that animal factors, in particular rumen fractional outflow rate (FOR) and rumen volume, have an influence on the between-sheep variation in methane (CH4) emission. Sheep were fed 2-hourly on chaffed lucerne hay. Following an acclimatization period of 21 days, the experiment lasted 16 days. Energy and nitrogen (N) balances were measured on days 1-6. Cr-EDTA marker was continuously infused into the rumen from day 9 to 16, and rumen contents emptied and sampled on days 13 and 16. Particulate and fluid FOR were estimated using feed lignin and Cr-EDTA, respectively. Daily CH, production was measured by the sulphur hexafluoride tracer technique on days 2, 5, 6, 12 and 15 of the experiment. CH4 production (g/day) was positively correlated with the pool size of organic matter (OM) in the rumen (OM pool, g) (r = 0.84, P = 0.002), OM intake (OMI, g/day) (r = 0.67, P = 0.04), and the rumen fill (g. wet digesta) (r = 0.76, P = 0.01). Multiple regression analysis showed that CH4 production was best predicted (R-2 = 0.88) as a function of OM pool and the molar % of butyrate; however, OM pool alone accounted for a large proportion (R-2 = 0.71) of the variation in CH4 production. CH4 yield (% gross energy intake, % GEI) was negatively correlated with the particulate FOR (%/h) ( r= -0.75, P = 0.01) and buffering capacity of rumen fluid (mmol HCl) (r = -0.72, P = 0.02) but positively correlated with the digestibility of cellulose (r = 0.66, P = 0.04). Multiple regression analysis showed that CH, yield was best predicted as a function of particulate FOR, OMI (g/kg liveweight(0.75)) and the molar % of butyrate (R-2 = 0.88). Particulate FOR alone explained a large proportion (R-2 = 0.57) of the variation in CH4 Yield. Particulate FOR was negatively correlated with rumen fill (r = -0.69, P = 0.03) and digestibility of cellulose (r = -0.65, P = 0.04). These results suggest that sheep with lower rumen particulate FOR (i.e. longer rumen retention times) had larger rumen fills and higher fibre digestibilities and CH4 yields. If rumen particulate FOR is to be used as a tool for CH4 mitigation, the repeatability of its relationship to CH4 emission must be assessed, preferably under grazing conditions

Persistence of differences between sheep in methane emission under generous grazing conditions

Four low and four high methane (CH4) emitters were selected from a flock of 20 Romney sheep on the basis of CH4 production rates per unit of intake, measured at grazing using the sulphur hexafluoride (SF,) tracer technique. Methane emissions from these sheep were monitored at grazing for four periods (P): October, November, January and February 1999/2000. All measurements were carried out on perennial ryegrass/white clover pasture under generous herbage allowance, and the sheep were maintained on similar pastures during non-measurement periods. The tracer technique was used for all the CH4 measurements and feed DM intake was calculated from total faecal collection and estimated DM digestibility. Data for liveweight (LW), gross energy intake (GEI) and CH4 emission were analysed using split-plot analysis of variance. In addition, a between-period rank order correlation analysis was carried out for CH4 emission data. Low CH4 emitters were heavier (P < 0.05) than the high emitters in all the periods, but they did not differ (P < 0.05) in their gross energy intakes (GEL MJ/kg LW0.75). Low and high CH4 emitters consistently maintained their initial rankings in CH4 yield (% GEI) throughout the subsequent periods and the correlation analysis of rank order for CH4 yield showed strong between-period correlation coefficients, although this was weaker in the last period. It is suggested that feeding conditions that maximize feed intake (e.g. generous allowance of good quality pasture under grazing) favour the expression and persistence of between-sheep differences in CH4 yield

Methane emission by alpaca and sheep fed on lucerne hay or grazed on pastures of perennial ryegrass/white clover or birdsfoot trefoil

Based on the knowledge that alpaca (Lama pacos) have a lower fractional outflow rate of feed particles (particulate FOR) from their forestomach than sheep (San Martin 1987), the current study measured methane (CH4) production and other digestion parameters in these species in three successive experiments (1, 2 and 3): Experiment 1, lucerne hay fed indoors; Experiment 2, grazed on perennial ryegrass/white clover pasture (PRG/WC); and Experiment 3, grazed on birdsfoot trefoil (Lotus corniculatits) pasture (Lotus). Six male alpaca and six castrated Romney sheep were simultaneously and successively fed on the forages either ad libitium or at generous herbage allowances (grazing). CH4 production (g/day) (using the sulphur hexafluoride tracer technique), voluntary feed intake (VFI), diet quality, and protozoa counts and volatile fatty acid concentrations in samples of forestomach contents were determined. In addition, feed digestibility, energy and nitrogen (N) balances and microbial N supply from the forestomach (using purine derivatives excretion) were measured in Experiment 1. Diets selected by alpaca were of lower quality than those selected by sheep, and the voluntary gross energy intakes (GEI, MJ) per kg of liveweight(0.75) were consistently lower (P0.05) in their CH4 yields (% GEI) when fed on lucerne hay (5.1 v. 4.7), but alpaca had a higher CH4 yield when fed on PRG/WC (9.4 v. 7.5, P0.05) in diet N partition or microbial N yield, but alpaca had higher (P<0.05) neutral detergent fibre digestibility (0.478 v. 0.461) and lower (P<0.01) urinary energy losses (5.2 v. 5.8 % GEI) than sheep. It is suggested that differences between these species in forestomach particulate FOR might have been the underlying physiological mechanism responsible for the differences in CH4 yield, although the between-species differences in VFI and diet quality also had a major effect on it

Methane emissions from pastoral systems: the situation in New Zealand

Methane is the major greenhouse gas of concern to countries like New Zealand and Uruguay, which have large ruminant and small human populations. The paper reviews the major factors affecting methane emission from ruminants. The relationship between DM intake and methane emission (g/d) is positive, but not strong. However, there is a stronger, but negative correlation when methane emission per unit of feed intake (kJ/100 kJ) is plotted against DM intake, suggesting that as intake increases the percentage of dietary energy lost as methane decreases. Starch and lipid are negatively correlated and fibre positively correlated with methane emission. The relationship between digestibility and methane is confounded with the effects of feed intake level: at low intakes methane increases as digestibility increases, but at high intakes methane decreases as digestibility increases. The SF6 tracer technique for estimating the methane emission of individual grazing animals is described and evaluated. Provided care is taken with SF6 permeation tube calibration it is considered that the technique gives reliable and repeatable estimates. Possible methane mitigation strategies are discussed including: reducing livestock numbers, increasing the efficiency of livestock production, exploiting natural between-animal variation in methane emission, dietary chemical additives that reduce methane, immunisation and manipulation of the rumen microbial ecosystem. While there are many interesting possibilities, more research is required before any of them is likely to be economically feasible. Methane mitigation strategies being developed in New Zealand to meet its obligations to the Kyoto Protocol are discussed. Research is focussed on developing accurate inventory methodology based on field measurements of various livestock classes and pasture types, and on possible mitigation technologies such as evaluating the cause of between-animal differences, plant inhibitors, and manipulation of the rumen microbial ecosystem

Marker concentration patterns of labelled leaf and stem particles in the rumen of cattle grazing bermuda grass (Cynodon dactylon) analysed by reference to a raft model

Large (>1600 mum), ingestively masticated particles of bermuda grass (Cynodon dactylon L. Pers.) leaf and stem labelled with Yb-169 and Ce-144 respectively were inserted into the rumen digesta raft of heifers grazing bermuda grass. The concentration of markers in digesta sampled from the raft and ventral rumen were monitored at regular intervals over approximately 144 h. The data from the two sampling sites were simultaneously fitted to two pool (raft and ventral rumen-reticulum) models with either reversible or sequential flow between the two pools. The sequential flow model fitted the data equally as well as the reversible flow model but the reversible flow model was used because of its greater application. The reversible flow model, hereafter called the raft model, had the following features: a relatively slow age-dependent transfer rate from the raft (means for a gamma 2 distributed rate parameter for leaf 0.0740 v. stem 0.0478 h(-1)), a very slow first order reversible flow from the ventral rumen to the raft (mean for leaf and stem 0.010 h(-1)) and a very rapid first order exit from the ventral rumen (mean of leaf and stem 0.44 h(-1)). The raft was calculated to occupy approximately 0.82 total rumen DM of the raft and ventral rumen pools. Fitting a sequential two pool model or a single exponential model individually to values from each of the two sampling sites yielded similar parameter values for both sites and faster rate parameters for leaf as compared with stem, in agreement with the raft model. These results were interpreted as indicating that the raft forms a large relatively inert pool within the rumen. Particles generated within the raft have difficulty escaping but once into the ventral rumen pool they escape quickly with a low probability of return to the raft. It was concluded that the raft model gave a good interpretation of the data and emphasized escape from and movement within the raft as important components of the residence time of leaf and stem particles within the rumen digesta of cattle

Hypoxyprobe™ reveals dynamic spatial and temporal changes in hypoxia in a mouse model of endometrial breakdown and repair

BACKGROUND: Menstruation is the culmination of a cascade of events, triggered by the withdrawal of progesterone at the end of the menstrual cycle. Initiation of tissue destruction and endometrial shedding causes spiral arteriole constriction in the functional layer of the endometrium. Upregulation of genes involved in angiogenesis and immune cell recruitment, two processes that are essential to successful repair and remodelling of the endometrium, both thought to be induced by reduced oxygen has been reported. Evidence for stabilisation/increased expression of the transcriptional regulator hypoxia inducible factor in the human endometrium at menses has been published. The current literature debates whether hypoxia plays an essential role during menstrual repair, therefore this study aims to delineate a role for hypoxia using a sensitive detection method (the Hypoxyprobe™) in combination with an established mouse model of endometrial breakdown and repair. RESULTS: Using our mouse model of menses, during which documented breakdown and synchronous repair occurs in a 24 h timeframe, in combination with the Hypoxyprobe™ detection system, oxygen tensions within the uterus were measured. Immunostaining revealed striking spatial and temporal fluctuations in hypoxia during breakdown and showed that the epithelium is also exposed to hypoxic conditions during the repair phase. Furthermore, time-dependent changes in tissue hypoxia correlated with the regulation of mRNAs encoding for the angiogenic genes vascular endothelial growth factor and stromal derived factor (Cxcl12). CONCLUSIONS: Our findings are consistent with a role for focal hypoxia during endometrial breakdown in regulating gene expression during menses. These data have implications for treatment of endometrial pathologies such as heavy menstrual bleeding

Modelling of the effect of ELMs on fuel retention at the bulk W divertor of JET

Effect of ELMs on fuel retention at the bulk W target of JET ITER-Like Wall was studied with multi-scale calculations. Plasma input parameters were taken from ELMy H-mode plasma experiment. The energetic intra-ELM fuel particles get implanted and create near-surface defects up to depths of few tens of nm, which act as the main fuel trapping sites during ELMs. Clustering of implantation-induced vacancies were found to take place. The incoming flux of inter-ELM plasma particles increases the different filling levels of trapped fuel in defects. The temperature increase of the W target during the pulse increases the fuel detrapping rate. The inter-ELM fuel particle flux refills the partially emptied trapping sites and fills new sites. This leads to a competing effect on the retention and release rates of the implanted particles. At high temperatures the main retention appeared in larger vacancy clusters due to increased clustering rate

Tritium distributions on W-coated divertor tiles used in the third JET ITER-like wall campaign

Tritium (T) distributions on tungsten (W)-coated plasma-facing tiles used in the third ITER-like wall campaign (2015–2016) of the Joint European Torus (JET) were examined by means of an imaging plate technique and β-ray induced x-ray spectrometry, and they were compared with the distributions after the second (2013–2014) campaign. Strong enrichment of T in beryllium (Be) deposition layers was observed after the second campaign. In contrast, T distributions after the third campaign was more uniform though Be deposition layers were visually recognized. The one of the possible explanations is enhanced desorption of T from Be deposition layers due to higher tile temperatures caused by higher energy input in the third campaign

Modelling of tungsten erosion and deposition in the divertor of JET-ILW in comparison to experimental findings

The erosion, transport and deposition of tungsten in the outer divertor of JET-ILW has been studied for an HMode discharge with low frequency ELMs. For this specific case with an inter-ELM electron temperature at the strike point of about 20 eV, tungsten sputtering between ELMs is almost exclusively due to beryllium impurity and self-sputtering. However, during ELMs tungsten sputtering due to deuterium becomes important and even dominates. The amount of simulated local deposition of tungsten relative to the amount of sputtered tungsten in between ELMs is very high and reaches values of 99% for an electron density of 5E13 cm$^{-3}$ at the strike point and electron temperatures between 10 and 30 eV. Smaller deposition values are simulated with reduced electron density. The direction of the B-field significantly influences the local deposition and leads to a reduction if the E×B drift directs towards the scrape-off-layer. Also, the thermal force can reduce the tungsten deposition, however, an ion temperature gradient of about 0.1 eV/mm or larger is needed for a significant effect. The tungsten deposition simulated during ELMs reaches values of about 98% assuming ELM parameters according to free-streaming model. The measured WI emission profiles in between and within ELMs have been reproduced by the simulation. The contribution to the overall net tungsten erosion during ELMs is about 5 times larger than the one in between ELMs for the studied case. However, this is due to the rather low electron temperature in between ELMs, which leads to deuterium impact energies below the sputtering threshold for tungsten