Effect of the application of cattle urine with or without the nitrification inhibitor DCD, and dung on greenhouse gas emissions from a UK grassland soil.

Emissions of nitrous oxide (N2O) from soils from grazed grasslands have large uncertainty due to the great spatial variability of excreta deposition, resulting in heterogeneous distribution of nutrients. The contribution of urine to the labile N pool, much larger than that from dung, is likely to be a major source of emissions so efforts to determine N2O emission factors (EFs) from urine and dung deposition are required to improve the inventory of greenhouse gases from agriculture. We investigated the effect of the application of cattle urine and dung at different times of the grazing season on N2O emissions from a grassland clay loam soil. Methane emissions were also quantified. We assessed the effect of a nitrification inhibitor, dicyandiamide (DCD), on N2O emissions from urine application and also included an artificial urine treatment. There were significant differences in N2O EFs between treatments in the spring (largest from urine and lowest from dung) but not in the summer and autumn applications. We also found that there was a significant effect of season (largest in spring) but not of treatment on the N2O EFs. The resulting EF values were 2.96, 0.56 and 0.11% of applied N for urine for spring, summer and autumn applications, respectively. The N2O EF values for dung were 0.14, 0.39 and 0.10% for spring, summer and autumn applications, respectively. The inhibitor was effective in reducing N2O emissions for the spring application only. Methane emissions were larger from the dung application but there were no significant differences between treatments across season of application.

Measurement at the field scale of soil delta13C and delta15N under improved grassland.

Variations in natural abundance of carbon (C) and nitrogen (N) stable isotopes are widely used as tools for many aspects of scientific research. By examining variations in the ratios of heavy to light stable isotopes, information can be obtained as to what physical, chemical and biological processes may be occurring. The spatial heterogeneity of soil delta(15)N- and delta(13)C-values across a range of scales and under different land use have been described by a number of researchers and the natural abundances of the C and N stable isotopes in soils have been found to be correlated with many factors including hydrology, topography, land use, vegetation cover and climate. In this study the Latin square sampling +1 (LSS+1) sampling method was compared with a simple grid sampling approach for delta(13)C and delta(15)N measurement at the field scale. A set of 144 samples was collected and analysed for delta(15)N and delta(13)C from a 12 x 12 grid (in a 1 ha improved grassland field in south-west England). The dimension of each cell of the grid was approximately 11 x 6 m. The 12 x 12 grid was divided into four 6 x 6 grids and the LSS+1 sampling technique was applied to these and the main 12 x 12 grid for a comparison of sample means and variation. The LSS+1 means from the 12 x 12 grid and the four 6 x 6 grids compared well with the overall grid mean because of the low variation within the field. The LSS+1 strategy (13 samples) generated representative samples from the 12 x 12 grid, and hence would be an acceptable method for sampling similar plots for the measurement of mean isotopic composition.

The contribution of cattle urine and dung to nitrous oxide emissions: Quantification of country specific emission factors and implications for national inventories.

Urine patches and dung pats from grazing livestock create hotspots for production and emission of the greenhouse gas, nitrous oxide (N2O), and represent a large proportion of total N2O emissions in many national agricultural greenhouse gas inventories. As such, there is much interest in developing country specific N2O emission factors (EFs) for excretal nitrogen (EF3, pasture, range and paddock) deposited during gazing. The aims of this study were to generate separate N2O emissions data for cattle derived urine and dung, to provide an evidence base for the generation of a country specific EF for the UK from this nitrogen source. The experiments were also designed to determine the effects of site and timing of application on emissions, and the efficacy of the nitrification inhibitor, dicyandiamide (DCD) on N2O losses. This co-ordinated set of 15 plot-scale, year-long field experiments using static chambers was conducted at five grassland sites, typical of the soil and climatic zones of grazed grassland in the UK. We show that the average urine and dung N2O EFs were 0.69% and 0.19%, respectively, resulting in a combined excretal N2O EF (EF3), of 0.49%, which is <25% of the IPCC default EF3 for excretal returns from grazing cattle. Regression analysis suggests that urine N2O EFs were controlled more by composition than was the case for dung, whilst dung N2O EFs were more related to soil and environmental factors. The urine N2O EF was significantly greater from the site in SW England, and significantly greater from the early grazing season urine application than later applications. Dycandiamide reduced the N2O EF from urine patches by an average of 46%. The significantly lower excretal EF3 than the IPCC default has implications for the UK's national inventory and for subsequent carbon footprinting of UK ruminant livestock products.

Fatty acid profiles associated with microbial colonization of freshly ingested grass and rumen biohydrogenation.

Two in situ studies were conducted to examine the use of odd-chain fatty acid profiles to study microbial colonization of freshly ingested herbage in the rumen as well as fatty acid biohydrogenation. In the first study, fresh perennial ryegrass was subjected to a range of sample preparation methods before incubation in the rumen for 2 or 7 h. In the second study, fresh perennial ryegrass was chopped into 1-cm lengths and incubated in polyester bags in the rumen for 2, 8, and 24 h. After removal of bags from the rumen, 4 different washing methods, ranging from manual squeezing to machine washing, were applied. Fatty acids were extracted from washed residues and determined, as methyl esters, by gas chromatography. The main odd-chain fatty acids (with the exception of anteiso C(15:0)) were not found in fresh grass and were useful markers of the effects of incubation time, sample preparation method, and washing method on microbial colonization/contamination. The concentration of these and other odd-chain fatty acids increased with incubation time in both studies. The results indicate rapid and continued microbial colonization of freshly ingested forages, although patterns of odd-chain fatty acids did not reveal any further information about the types of bacteria-colonizing herbage. Principal component, biplot analysis provided a useful overall description of the processes of microbial colonization and degradation of plant fatty acids on fresh herbage incubated in the rumen. Bolus formation during mastication and ingestion results in extensive damage to herbage; none of the techniques (cutting, crushing, and drying/grinding) investigated in this work was able to replicate the effects of bolus formation in the animal. The study provided further evidence of loss of unfermented feed particles through polyester bag pores, especially when feeds are dried and ground. Biohydrogenation of the polyunsaturated fatty acids of fresh herbage was used principally by solid-associated bacteria to enable them to take up high levels of trans-11 C(18:1) and C(18:0) fatty acids. Although trans-11 C(18:1) was strongly associated with bacterial markers (odd- and branched-chain fatty acids), its precursor (cis-9, trans-11 C(18:2)) was not associated with bacterial variation, suggesting that its production in the rumen under these conditions was mainly extracellular.

APICAL INTRUSIVE GROWTH OF FIBRES FROM THE FLOWERING STEM OF LOLIUM TEMULENTUM L.

Extension of the internode below the point of insertion of the flag leaf of Lolium temulentum L. takes place in the lower 10 to 15 mm. Extension of the long epidermal cells also takes place in this region. However, fibre cells show little extension in the lower 10 mm and maximum extension occurs at about 30 mm above the node. Thus, there is extension of fibre cells within that portion of the internode which has ceased extension and is bounded by an epidermis, the cells of which have also ceased elongation. This suggests the occurrence of intrusive growth of the fibres and evidence for this is shown from the measurement of macerated fibres and from electron micrographs. The walls of fibre cells measured in internodes of different lengths and ages thicken only very slightly in the lower 30 mm and most cell wall thickening occurs between 45 and 90 mm above the node. Thus most cell wall thickening occurs after the fibre cells have ceased to elongate.

Statistical evaluation of mathematical models for microbial growth.

The aim of this study was to evaluate the suitability of several mathematical functions for describing microbial growth curves. The nonlinear functions used were: three-phase linear, logistic, Gompertz, Von Bertalanffy, Richards, Morgan, Weibull, France and Baranyi. Two data sets were used, one comprising 21 growth curves of different bacterial and fungal species in which growth was expressed as optical density units, and one comprising 34 curves of colony forming units counted on plates of Yersinia enterocolitica grown under different conditions of pH, temperature and CO(2) (time-constant conditions for each culture). For both sets, curves were selected to provide a wide variety of shapes with different growth rates and lag times. Statistical criteria used to evaluate model performance were analysis of residuals (residual distribution, bias factor and serial correlation) and goodness-of-fit (residual mean square, accuracy factor, extra residual variance F-test, and Akaike's information criterion). The models showing the best overall performance were the Baranyi, three-phase linear, Richards and Weibull models. The goodness-of-fit attained with other models can be considered acceptable, but not as good as that reached with the best four models. Overall, the Baranyi model showed the best behaviour for the growth curves studied according to a variety of criteria. The Richards model was the best-fitting optical density data, whereas the three-phase linear showed some limitations when fitting these curves, despite its consistent performance when fitting plate counts. Our results indicate that the common use of the Gompertz model to describe microbial growth should be reconsidered critically, as the Baranyi, three-phase linear, Richards and Weibull models showed a significantly superior ability to fit experimental data than the extensively used Gompertz.

Comparison of mathematical models to describe disappearance curves obtained using the polyester bag technique for incubating feeds in the rumen.

Different nonlinear models were evaluated as candidates to describe ruminal degradation kinetics of forages from data obtained by the incubation of the feeds in the rumen using polyester bags. Nine models were used: segmented model with three straight lines (Mod0); simple Mitscherlich or exponential (Mod1); inverse polynomial (Mod2); compartmental model with two exponential terms (Mod3); generalized Mitscherlich (Mod4); generalized Michaelis-Menten (Mod5); logistic (Mod6); Gompertz (Mod7); and generalized Von Bertalanffy (Mod8). All these models can be represented in the general form D = W + S0 x phi(t), where D is in situ disappearance at incubation time t, W and S0 are positive scalars, and phi is a positive monotonically increasing function unique to each of the models studied. Based on first principles, a general formula for calculating the extent of degradation of feeds in the rumen has been derived that is applicable to all the models. The disappearance curves of different feed components (DM, N, and NDF) of 87 Mediterranean forages (i.e., a total of 261 curves) were fitted to all the models. A comparative study was carried out based on the mathematical, statistical, and biological characteristics of the models. Flexible models that can accommodate both diminishing returns and sigmoidal behavior were more appropriate in describing the curves. A discrete-lag parameter was introduced into Mod0, Mod1, and Mod2 to describe the initial stage of the disappearance curve, and this parameter considerably improved the fit of experimental data. Based on statistical criteria, models Mod1, Mod4, Mod5, and Mod8 were better than the others for most statistical tests and disappearance curves, but differences among these four models were not consistent. The estimates of degradation parameters to quantify the rate (half-life, fractional degradation rate), and extent (undegradable fraction, effective degradability) of ruminal degradation of feeds were also used as a means to discriminate between models, although in most cases all of the models gave similar values of the degradation parameters. In particular, when the extent of degradation was calculated for each forage and feed component, differences between the estimates obtained with the different models were of little nutritional significance for the animal.

A generalized Michaelis-Menten equation for the analysis of growth.

The functional form W = (W0Kc + Wf t(c)) /(Kc + t(c)), where W is body size at age t, W0 and Wf are the zero- and infinite-time values of W, respectively, and K and c are constants, is derived. This new generalized Michaelis-Menten-type equation provides a flexible model for animal growth capable of describing sigmoidal and diminishing returns behavior. The parameters of the nonlinear model are open to biological interpretation and can be used to calculate reliable estimates of growth traits, such as maximum or average postnatal growth rates. To evaluate the new model, the derived equation and standard growth functions such as the Gompertz and Richards were used to fit 83 growth data sets of different animal species (fish, mice, hamsters, rats, guinea pigs, rabbits, cats, dogs, broilers, turkeys, sheep, goats, pigs, horses, and cattle) with a large range in body size. A comparative study was carried out based on mathematical, statistical, and biological characteristics of the models. The statistical goodness-of-fit achieved with the new model was similar to that of Richards, and both were slightly superior to the Gompertz. The new model differed from the others with respect to some of the estimated growth traits, but there were highly significant correlation coefficients between estimates obtained with the different models, and the ranking of animals based on growth parameters computed with the new function agreed with the rankings computed by the other models. Therefore, the new model, with its variable inflection point, was able to adequately describe growth in a wide variety of animals, to fit a range of data showing sigmoidal growth patterns, and to provide satisfactory estimates of traits for quantifying the growth characteristics of each type of animal.

Technical note: A proposed method to determine the extent of degradation of a feed in the rumen from the degradation profile obtained with the in vitro gas production technique using feces as the inoculum.

A method is proposed to determine the extent of degradation in the rumen involving a two-stage mathematical modeling process. In the first stage, a statistical model shifts (or maps) the gas accumulation profile obtained using a fecal inoculum to a ruminal gas profile. Then, a kinetic model determines the extent of degradation in the rumen from the shifted profile. The kinetic model is presented as a generalized mathematical function, allowing any one of a number of alternative equation forms to be selected. This method might allow the gas production technique to become an approach for determining extent of degradation in the rumen, decreasing the need for surgically modified animals while still maintaining the link with the animal. Further research is needed before the proposed methodology can be used as a standard method across a range of feeds.

Correcting the calculation of extent of degradation to account for particulate matter loss at zero time when applying the polyester bag method.

The established calculation of ruminal extent of degradation using the polyester bag method overestimates extent. The wash fraction, at least in part, is subject to losses from the rumen due to passage. Four formulae are proposed to minimize this risk of overestimation. Four options are considered: 1) passage losses for particulate matter escaping from the bag at zero time are according to the particulate fractional passage rate, 2) the liquid rate, and 3) the average of the two, and additionally 4) there is no instantly degradable fraction. The established and proposed formulae were examined using polyester bag data for six forages: grass silage, fresh ryegrass, fresh white clover, alfalfa hay, mixed grasses hay, and hay from a permanent mountain meadow. The established formula gave appreciably higher estimates of extent of degradation in all cases. Overestimation was at least 6.9, 4.9, 2.9, 2.1, 2.4, and 4.5%, respectively, for the six forages.

Alternative approaches to predicting methane emissions from dairy cows.

Previous attempts to apply statistical models, which correlate nutrient intake with methane production, have been of limited value where predictions are obtained for nutrient intakes and diet types outside those used in model construction. Dynamic mechanistic models have proved more suitable for extrapolation, but they remain computationally expensive and are not applied easily in practical situations. The first objective of this research focused on employing conventional techniques to generate statistical models of methane production appropriate to United Kingdom dairy systems. The second objective was to evaluate these models and a model published previously using both United Kingdom and North American data sets. Thirdly, nonlinear models were considered as alternatives to the conventional linear regressions. The United Kingdom calorimetry data used to construct the linear models also were used to develop the three nonlinear alternatives that were all of modified Mitscherlich (monomolecular) form. Of the linear models tested, an equation from the literature proved most reliable across the full range of evaluation data (root mean square prediction error = 21.3%). However, the Mitscherlich models demonstrated the greatest degree of adaptability across diet types and intake level. The most successful model for simulating the independent data was a modified Mitscherlich equation with the steepness parameter set to represent dietary starch-to-ADF ratio (root mean square prediction error = 20.6%). However, when such data were unavailable, simpler Mitscherlich forms relating dry matter or metabolizable energy intake to methane production remained better alternatives relative to their linear counterparts.

Aspects of the metabolism of dairy cows during the incubation of bovine spongiform encephalopathy.

As part of a nutritional study lasting from six weeks before calving to 22 weeks of lactation, blood samples collected from 47 dairy cows maintained under well-defined conditions were analysed for a variety of metabolites and hormones. At various times after the completion of the study, six of the animals developed clinical signs of bovine spongiform encephalopathy (BSE), although when they were sampled it was not known that they were incubating the disease. The data were used to make comparisons between the animals that developed BSE and those that did not develop the disease and which had been maintained under the same conditions. The greatest differences between the animals incubating BSE and the control animals were observed at times of nutritional stress, at the start of lactation and when the intake of concentrate feeds was reduced at week 13 of lactation. In the animals that subsequently developed BSE, feed intakes were lower in early lactation; plasma beta-hydroxbutyrate concentrations were significantly higher (P<0.001) at weeks 3 and 5 of lactation; adjusted milk yields were lower until week 6 of lactation and milk fat concentrations were consistently lower. There was no effect on plasma glucose concentrations, although insulin concentrations were significantly lower in week 1 of lactation (2-27 v 2.50 microiu/ml) (P<0.05). The concentrations of plasma proteins and urea were unaffected by BSE incubation, apart from protein concentrations being significantly higher one week before calving, and the concentration of urea being significantly lower five weeks before calving. The plasma concentrations of somatotrophin, prolactin, oestradiol and progesterone were similar in the two groups of animals throughout the study. The differences observed indicate that the energy metabolism of dairy cows incubating BSE may be subtly altered before the onset of clinical signs of the disease.

A simple but flexible growth function.

The functional form W(t) = Wf - (Wf - Wo) exp[-k infinite (t - T) + 2c(root of t - root of T)] where Wf, Wo, k infinite, c and T are constants, is derived as a growth equation and evaluated using commonly applied growth functions such as the Gompertz, logistic, monomolecular and Richards. Further evaluation is made with reference to sets of observations on growth in a number of animal species ranging from mice to horses. The new function provides a flexible growth equation capable of describing sigmoidal and diminishing returns behaviour. It appears adept at describing sigmoidal patterns exhibiting faster early growth and a fairly low but variable point of inflexion, and can therefore be perceived as a generalised Gompertz equation. The function also has the ability to describe a wide range of hyperbolic shapes when there is no point of inflexion. The analysis described suggests that this simple equation is a worthwhile addition to the corpus of growth functions.

Adaptation of compartmental schemes for interpreting isotope dilution data on volatile fatty acid metabolism in the rumen to the non-steady state and for single-dose injection.

A unifying mathematical analysis of the one-pool and three-pool schemes for estimating acetate, propionate and butyrate production in the rumen from isotope dilution data is presented. Emphasis is given to non-steady-state conditions and to the administration of isotope as a single injection. The single-pool representation is also derived as a special case of the three-pool representation by assuming that the external fluxes are directly proportional to their concentrations in the rumen.

Studies of method of conserving grass herbage and frequency of feeding in cattle. 1. Voluntary feed intake, digestion and rate of passage.

The effect of method of conserving grass herbage and the frequency of feeding on digestion in and passage from the rumen was studied in growing cattle. A single sward of perennial ryegrass (Lolium perenne cv. Endura) was cut on 1 d and conserved as silage or hay. These forages were offered as the only feed to twelve rumen-cannulated Friesian steers (average initial live weight (LW) 128 kg) either at a restricted level of intake (20 g dry matter (DM)/kg LW; Expt 1) or ad lib. (Expt 2). In Expt 1 the forages were offered either once or eight times daily. When fed once or eight times daily, in vivo digestibility of DM was very similar for hay and silage (0.771 and 0.783 respectively), while the fractional rate of digestion measured by incubation in Dacron bags was significantly (P less than 0.05) higher for silage (0.069) than hay (0.057). The rate of passage of mordanted feed out of the rumen was significantly (P less than 0.01) faster for silage when determined from concentrations of marker in the rumen (0.034 v. 0.028/h). The volume of liquid in the rumen was lower (P less than 0.01) for animals consuming silage (27.51) compared with hay (37.11), while the fractional rate of passage of liquid from the rumen was higher (0.141 v. 0.098/h, P less than 0.05). The volume of liquid in the rumen was also lower (P less than 0.05) in steers offered feed eight times daily (29.5 l), compared with once daily (35.1 l), but frequency of feeding had no significant effect either on total digestibility or on rate of digestion in the rumen. The ad lib. intake of silage was significantly (P less than 0.01) lower than that of hay (4.53 v. 5.16 kg DM/d) in Expt 2.

On using a double-exponential model for describing faecal marker concentration curves.

A mathematical and numerical appraisal of the double-exponential model is presented, with particular reference to its ability to reproduce preset values of biological measures such as rumen and whole-tract mean retention times and faecal production rate. A complete, formal mathematical derivation is provided, with consideration given to a solution to the model when the two smallest rate constants are equal. It is concluded that any error introduced by using an approximation in deriving the double-exponential form is negligible in practice, and that the double exponential in logarithmic form is able to determine the relevant biological measures quite satisfactorily and speedily.

A non-linear compartmental model to describe forage degradation kinetics during incubation in polyester bags in the rumen.

A new model for describing forage degradation kinetics during incubation in the rumen using polyester bags is presented. Attention is given to dealing with the problem of deviations from exponential behaviour in the early stages of degradation by devising a function capable of representing exponential or sigmoidal trends. This is achieved by allowing part of the fractional degradation rate to vary with time of incubation, thus enabling responses other than those expected under simple first-order kinetics to be described. Seven sets of data consisting of 620 curves were analysed to study the performance of the new model compared with a commonly used exponential model. The proportion of significantly better fits varied from set to set. The new model deals successfully with sigmoidal behaviour and, thus, provides a means of analysing the degradation profiles of low-quality forage feeds.

The effects of diffusion and viscosity on ruminant faecal excretion of markers in a simple compartmental model.

Using equations obtained in a previous analysis, results are computed numerically which illustrate the effects of diffusion and viscosity on faecal excretion patterns of markers in ruminants. Results are first given for plug flow, a velocity gradient produced by viscosity, and diffusion, each mechanism operating alone. Plug flow gives a period during which no marker appears in the faeces, then a sharp increase in faeces marker concentration, followed by rapid washout. A velocity gradient gives a more gradual appearance of marker in the faeces followed by a slower washout. Diffusion alone (although not realistic for ruminant marker kinetics) can give early appearance of marker followed by slow washout. Combining diffusion with a velocity gradient produced by viscosity can give a range of behaviour, depending on the effective diffusion coefficient, D'; an approximate method is used to compute these solutions. Because plug flow with no velocity gradient plus diffusion gives results similar to convective flow with a velocity gradient plus diffusion, we believe it will not be possible to determine the main mechanisms defining marker outflow patterns from observations of marker kinetics alone, and more detailed investigations will be needed. Although estimates of quantities such as mean transit time are unaffected by detailed mechanism, the interpretations of measures such as sigmoidicity, sharpness of the faecal marker concentration v. time curve, and length and nature of the washout tail are highly dependent on mechanism.

A unifying mathematical analysis of methods to estimate rumen volume using digesta markers and intraruminal sampling.

A theoretical exposition of methods for estimating the quantity of digesta in the rumen using digesta-flow markers and intraruminal sampling is given, with emphasis on the kinetic assumptions underlying each method. Single- and dual-marker approaches to estimating volume in steady and non-steady-state are presented.

Rumen fill and digesta kinetics in lactating Friesian cows given two levels of concentrates with two types of grass silage ad lib.

Four lactating Friesian cows with permanent cannulas in the rumen and proximal duodenum were given early (EC)- or late (LC)-cut grass silage ad lib., each with either 3 or 9 kg concentrate dry matter (DM)/d in a 4 x 4 Latin square design starting about 10 weeks after calving. Feed was offered twice daily at 08.30 hours and 15.30 hours. Periods lasted 5 weeks and measurements were made in the last 15 d. The higher amount of concentrates increased total DM intake but reduced silage DM intake and the fractional rate of degradation of silage-fibre DM. Later cutting date of silage had no effect on DM intake or the fractional rate of degradation of silage-fibre DM but reduced potential degradability of silage fibre. Dilution rate of CoEDTA in rumen fluid was greater during the day (eating period) than during the night (resting period). Dilution rates measured at the duodenum were lower than those measured in the rumen, but neither was affected by diet. Silage-particle passage rates were measured by use of ytterbium-labelled silage fibre (Yb-fibre) and chromium-mordanted faecal particles (Cr-faeces) and samples were taken at the duodenum and in the faeces. Values for slower rate constant (k1) and transit time were higher and for faster rate constant (k2) were lower for Yb-fibre than for Cr-faeces, but differences in total mean retention time were inconsistent. Values for k1 for both markers and k2 for Yb-fibre only were similar at both sampling sites, but values for k2 for Cr-faeces were lower in the faeces. No diet effects were established with Yb-fibre but, with Cr-faeces, k1 was reduced by more concentrates and EC-silage. Daily mean weights of wet digesta, liquid, neutral-detergent fibre (NDF) and indigestible NDF in the rumen were greater with LC-silage but were unaffected by the amount of concentrates whereas weight of rumen DM was increased by the amount of concentrates only. Maximum rumen fill occurred at 18.00 hours with all diets. Particle-size distribution of rumen contents did not vary markedly during the day. Mean particle size was generally greater with LC-silage than EC-silage. Very small particles, passing through the 0.3 mm screen, constituted about half the rumen DM. It is concluded that rumen fill could have limited intake of LC-silage but not EC-silage.(ABSTRACT TRUNCATED AT 400 WORDS)