The development of an intraruminal nylon bag technique using non-fistulated animals to assess the rumen degradability of dietary plant materials.

Although the conventional in situ ruminal degradability method is a relevant tool to describe the nutritional value of ruminant feeds, its need for rumen-fistulated animals may impose a restriction on its use when considering animal welfare issues and cost. The aim of the present work was to develop a ruminal degradability technique which avoids using surgically prepared animals. The concept was to orally dose a series of porous bags containing the test feeds at different times before slaughter, when the bags would be removed from the rumen for degradation measurement. Bags, smaller than those used in the conventional nylon bag technique, were made from woven nylon fabric, following two shape designs (rectangular flat shape, tetrahedral shape) and were fitted with one of three types of device for preventing their regurgitation. These bags were used in two experiments with individually housed non-pregnant, non-lactating sheep, as host animals for the in situ ruminal incubation of forage substrates. The bags were closed at the top edge by machine stitching and wrapped in tissue paper before oral dosing. Standard times for ruminal incubation of substrates in all of the tests were 4, 8, 16, 24, 48, 72 and 96 h before slaughter. The purpose of the first experiment was to compare the effectiveness of the three anti-regurgitation device designs, constructed from nylon cable ties ('Z-shaped', ARD1; 'double Z-shaped', ARD2; 'umbrella-shaped', ARD3), and to observe whether viable degradation curves could be generated using grass hay as the substrate. In the second experiment, three other substrates (perennial ryegrass, red clover and barley straw) were compared using flat and tetrahedral bags fitted with type ARD1 anti-regurgitation devices. Non-linear mixed-effect regression models were used to fit asymptotic exponential curves of the percentage dry matter loss of the four substrates against time of incubation in the reticulorumen, and the effect of type of anti-regurgitation device and the shape of nylon bag. All three devices were highly successful at preventing regurgitation with 93% to 100% of dosed bags being recovered in the reticulorumen at slaughter. Ruminal degradation data obtained for tested forages were in accordance with those expected from the conventional degradability technique using fistulated animals, with no significant differences in the asymptotic values of degradation curves between bag shape or anti-regurgitation device. The results of this research demonstrate the potential for using a small bag technique with intact sheep to characterise the in situ ruminal degradability of roughages.

Feed intake and diet selection in Angus-cross heifers of two frame sizes at two stages of growth.

Assessing feed efficiency in pasture-based systems can be challenging due to difficulties in measuring feed intake and diet preference. Furthermore, heifers may modify their intake of a particular forage species depending on its nutritive and physical attributes and on their own physiological status. For instance, heifers growing to larger mature sizes have higher maintenance requirements, which may affect their dietary preferences. The present study was designed to determine the influence of frame size (FS) on feed intake and diet selection at 2 age subclasses: postweaning, at, on average, 313 d of age (317 kg BW for larger FS and 285 kg BW for smaller FS), and as yearlings, at, on average, 403 d of age (391 kg BW for larger FS and 343 kg BW for the smaller FS). Twenty-four Angus-cross heifers were evaluated at these ages. They were housed in a drylot equipped with a Broadbent Feeding System and had ad libitum access to cubed red clover and cubed fescue hay in separate buckets. Following 3-wk acclimation periods, DMI of each forage species was assessed daily for 10 d. Body weights were also recorded daily over those 10 d. Larger and smaller FS cattle did not differ in ADG, feed conversion ratio, and Kleiber ratio at either age (P > 0.05). However, heifers from larger FS category had higher total and red clover DMI at both ages (P < 0.05) and higher proportions of red clover in their diet as yearlings (P < 0.001). Cumulative differences between clover and fescue DMI were similar at postweaning in both FS (P > 0.05); however, at yearling age, the larger FS cattle ate consistently and cumulatively more red clover (P < 0.001). The intake and diet selection of heifers across ages were not consistent for either FS category (P = 0.748), suggesting difficulties in predicting lifetime feed efficiencies based on an early measure. Although gains and feed conversion ratios were similar between FS categories, the larger FS heifers ate more. At yearling age, these heifers of larger metabolic size also selected a greater proportion of red clover in their diet. Because the FS and, thereby, BW of cattle affect intakes and diet selections, they also may impact the composition and sustainability of grasslands generally. Therefore, the relationship between FS and pasture usage likely needs to be integrated into descriptions of cow efficiency in pasture-based operations.

Evaluation of long-chain alcohols as diet composition markers in goats grazing heathland areas.

An experiment was conducted to assess the potential of long-chain alcohols (LCOH), in alternative or combined with alkanes and long-chain fatty acids (LCFA), as faecal markers to estimate the diet composition of goats grazing heathland vegetation with associated improved pastures. A total of seven diets were offered across the grazing season. The diets were composed of mixtures of herb species (Lolium perenne and Trifolium repens) and woody species (Erica spp., Calluna vulgaris and Ulex gallii) in an attempt to simulate diet selection of goats on these complex vegetation areas. The diet composition was estimated using LCOH markers alone or combined with alkanes, LCFA and alkanes+LCFA, by least square optimization procedures. The data showed large differences between plant species in their LCOH profile. Generally, plant species showed higher LCOH concentrations than those of alkanes and lower than LCFA markers. Faecal recovery of LCOH was incomplete and increased in a linear manner (P < 0.001) with the carbon-chain length, and was influenced by diet composition and its digestibility. The diet composition estimates based on LCOH alone were more accurate (P < 0.05) than those using alkanes or LCFA alone. Results showed that the combination of LCOH with alkanes, LCFA and alkanes+LCFA resulted in more accurate (P < 0.05) estimates of diet composition, indicating that LCOH provided different discriminatory information to that of alkanes and LCFA, helping in the discrimination of the plant species used in this experiment. Results indicate that correction of faecal LCOH concentrations to incomplete faecal recovery is necessary to obtain more accurate estimates of diet composition. Nevertheless, it seemed that the use of a less accurate recovery correction on LCOH markers had a lower impact on the accuracy of estimates than that of alkanes and LCFA.

In vitro studies of the metabolism of [14C]- n-alkanes using ruminal fluid of sheep as substrate.

Whether the rumen microbes are able to synthesize and/or degrade long-chain alkanes in anaerobic conditions remains a question to be answered before these hydrocarbons can be confidently used as duodenal flow or rumen transit markers. In this context, an experiment in vitro was carried out to establish whether within a rumen liquor fermentation system, n-alkanes can be derived from de-waxed structures of the plant or from non-alkane wax components (long-chain fatty alcohols, long-chain fatty acids and esters), or may be metabolized by bacteria to other components or to shorter-chain hydrocarbons. Ryegrass was labelled with 14C in growth chambers under controlled conditions in order to use it as a substrate. The labelled material obtained was separated in three fractions: labelled alkanes, labelled de-waxed plant and labelled wax components without the alkanes. These fractions were used for three different incubations in vitro, which objectives were as follows: 1. To check whether rumen bacteria can synthesize alkanes from carbon structures other than waxes (e.g. sugars). 2. To verify whether rumen bacteria can metabolize the n-alkanes to other compounds. 3. To check whether rumen bacteria can synthesize n-alkanes from other carbon compounds from waxes. The results showed that there was neither bacterial synthesis nor metabolism of the n-alkanes in in vitro conditions.

The transfer of radiocaesium to ewes through a breeding cycle--an illustration of the pitfalls of the transfer coefficient.

A study to measure the transfer of radiocaesium to adult female sheep through a breeding cycle is described. The transfer of radiocaesium from the diet to muscle (estimated as the equilibrium transfer coefficient) was significantly lower to pregnant, and especially lactating, animals compared to non-lactating and barren animals. High dry matter intake rates were also associated with significantly lower transfer coefficients. Known relationships between dry matter intake rates and protein turnover could credibly explain some of these differences. However, when described as the concentration ratio, radiocaesium transfer to meat was apparently highest during lactation. The apparent difference in results obtained by the two approaches of determining transfer is the consequence of daily dry matter intake being a denominator within the estimation of transfer coefficient. A wider discussion of transfer coefficients and concentration ratios leads us to suggest that the concentration ratio is the more robust and potentially generic parameter.

The transfer of radionuclides from saltmarsh vegetation to sheep tissues and milk.

Radionuclides released into the Irish Sea by the Sellafield reprocessing plant are deposited onto tide-washed pastures along the western coast of the United Kingdom. Many of these pastures are grazed by sheep or cattle. This paper describes a controlled feeding study, in which saltmarsh vegetation harvested from close to the Sellafield plant, was fed to lambs and adult female sheep for a period of 8 weeks. Activity concentrations of (60)Co, (95)Nb, (106)Ru, (134)Cs, (137)Cs, (238)Pu, (239,240)Pu and (241)Am were determined in edible tissues and transfer parameters estimated. The activity concentrations of some of the radionuclides will not have been in equilibrium with those in the diet. Nevertheless, the study was reasonably realistic in terms of agricultural management as the period of the study was similar to that for which lambs graze on the saltmarshes. A field study to determine the activity concentrations of (137)Cs and (239,240)Pu in the milk of ewes grazing a saltmarsh close to Sellafield is also described.

The transfer of different forms of 35S to goat milk.

Sulphur-35 is released during the routine operation of UK gas-cooled reactors. An experiment to determine the rates of transfer of different forms of (35)S to goat milk is described. Lactating goats received (35)S orally as single administrations of sulphate, L-methionine, or grass contaminated either through root uptake of (35)S as sulphate or through aerial deposition of (35)S as carbonyl sulphide onto the grass. Transfer was higher for (35)S administered as methionine compared with (35)S administered as sulphate. Changes in activity concentrations in milk for all sources of (35)S demonstrated two components of loss. The first component had a half-life of circa 1 d for all sources, the second was longer in goats administered carbonyl sulphide (44 d) than in all of the other treatments (circa 10 d). The rate of transfer of (35)S to milk of a further group of goats receiving (35)S-sulphate daily appeared to reach equilibrium within 30 d. Extrapolation of transfer parameters derived to other dairy ruminants is discussed.

The transfer of 239/240Pu to cow milk.

In summer 1993 we measured the transfer of (239/240)Pu to milk from herbage from a pasture located 5 km from the Chernobyl nuclear power plant. In one treatment cows were allowed to graze freely on the pasture. In a second treatment, cows were fed herbage collected from the pasture in stalls. The milk transfer coefficient; F(m) did not vary significantly between treatments and the mean value of 7.5x10(-6)d l(-1) was higher than previously reported values. Despite many values of F(m) for Pu in the literature we identified few relevant original data sets. Transfer coefficient values for Pu are only appropriate when used in conjunction with a specified time period or an appropriate model which allows for the biological half-life. We recommend for screening purposes an F(m) value of 1x10(-5)d l(-1) for Pu, with an order of magnitude lower value being appropriate for cows which are only exposed for one grazing season.

Alcohol discovered in the urine after death: ante-mortem ingestion or post-mortem artefact?

A study was undertaken of blood and urine alcohol levels in 44 cases where the bodies had been immersed in water, following accidents, for at least one day after death. In no case was there any ante-mortem evidence of ingested alcohol. In 15 cases there was a raised urine alcohol. The average urine-to-blood alcohol ratio in these cases was 0.56:1. This is dramatically less that that seen after ingestion. It is suggested that the alcohol, following its initial post-mortem production in the abdomen, enters the urine by diffusion from surrounding tissues. The presence of alcohol in the urine from bodies that have not been recovered and examined until several days after death cannot always be taken to indicate ante-mortem ingestion. Urine-to-blood alcohol ratios of less than 1:1 are strongly suggestive of post-mortem production.

A simple method for the estimation of the bioavailability of radiocaesium from herbage contaminated by adherent soil.

Adherent soil may contribute a large proportion of the radiocaesium content of sampled vegetation. Consequently, inadvertent ingestion of adherent soil can contribute significantly to the radiocaesium intake of grazing animals, and needs to be accounted for within radiological assessments. However, accurate estimation of the degree of soil adhesion on vegetation is acknowledged to be difficult. To determine the relative contributions of vegetation and soil to the radiocaesium contamination of milk and tissues, soil-specific estimation of radiocaesium bioavailability values would be required. Here we suggest that a previously developed in-vitro bioavailability assay (involving a 2 h extraction with 0.1 M stable CsCl) can be used to estimate the true absorption coefficient of radiocaesium associated with sampled vegetation directly. Using this technique, seasonal trends in bioavailability are demonstrated to vary in accordance with estimations of the degree of soil adherent to vegetation collected from an upland pasture. The use of this technique would negate the need for detailed measurements of the amount of soil adhering to sampled vegetation and soil-specific radiocaesium bioavailability assessments.

Radiocaesium variability within sheep flocks: determination of 137Cs intake in free-ranging sheep.

A study is described in which 137Cs intake by free-ranging sheep was estimated at two farms in the area of west Cumbria (northwest England) which received some of the highest amounts of fallout from the Chernobyl accident within the United Kingdom. The faecal excretion of 137Cs was estimated from faecal 137Cs activity concentrations and the use of intraruminal controlled release devices containing Cr2O3 to determine faecal dry matter output. The intake of 137Cs was estimated by assuming an apparent absorption coefficient appropriate to the herbage grazed. The methodology has the advantage that sampling of herbage representative of that ingested by study animals is not required. Caesium-137 dietary intake explained >60% of the observed variability in the 137Cs activity concentration determined in the muscle of sheep. Resultant transfer coefficient (Ff) values to describe the transfer of 137Cs from the diet to muscle were in agreement with previously reported values. At one farm, there was a positive correlation between the 137Cs activity concentration in muscle and Ff whilst at the other farm there was a negative correlation between Ff and 137Cs dietary intake. Potential reasons for these observations are discussed.

A model of radioiodine transfer to goat milk incorporating the influence of stable iodine.

Previously reported models for radioiodine in ruminants cannot account for the effect of variations in stable iodine intake including large countermeasure doses of stable iodine on the transfer of radioiodine to goat milk. A metabolically based model of radioiodine transfer in goats has been parameterised using new experimental data on the effect of countermeasure doses of stable iodine on radioiodine transfer to milk. To account for the effect of dietary stable iodine levels, the model represents the transfer of iodine from the extracellular fluid to milk with Michaelis-Menten kinetics. The model shows good agreement with the experimental data, and the estimated parameters compare favourably with values which can be estimated from the literature. The parameterised model accounts for 95% of the variation in the observed data for milk, faeces, urine and thyroid (n=199). The model has been used to predict the effects of variation in stable iodine intake and the extent of consequent chemical contamination of milk by stable iodine. The time taken for radio-iodine to reach peak concentrations in milk following a deposition event is predicted to vary significantly (ca. 2 days) over a range of expected stable iodine intakes. Doses of stable iodine sufficient to reduce the radioiodine transfer to milk will result in stable iodine concentrations in milk greatly in excess of internationally advised limits. Therefore, we recommend that stable iodine supplementation not be used as a countermeasure to reduce radioiodine transfer to milk. Indeed, model predictions suggest that reductions in stable iodine intake would be a more effective countermeasure. However, this is unlikely to be feasible since the short physical half-life of 131I may not allow adequate time to implement changes in feed manufacture. The model described in this paper is freely available in ModelMaker 3.0 format (http://www.notingham.ac.uk/environmentalmodelling+ ++/).

Measurement of dietary nutrient intake in free-ranging mammalian herbivores.

The nutrient intakes of mammalian herbivores depend on the amount and the nutrient content of the plant species and plant parts which they eat. We review the merits of oesophageal-fistulated (OF) animals, microhistological procedures, stable C-isotope discrimination and plant cuticular-wax markers as methods for estimating diet composition and intake in both ruminant and non-ruminant herbivores. We also briefly discuss methods based on grazing behaviour measurements or on H2O or Na turnover, and methods for estimating supplement or soil intake. Estimates of intake in ruminants are often based on separate measurements of faecal output and herbage digestibility. We review this approach and emphasize that, under some circumstances, the applicability of in vitro digestibility estimates based on OF extrusa is questionable. We discuss how plant-wax marker patterns can be used to check whether OF and test animals are consuming similar diets, but also emphasize that a major advantage of the use of plant-wax markers is that this approach may obviate altogether the need for OF animals. Estimates of total herbage intake can be partitioned into the intakes coming from different plant species and/or parts, provided diet composition can be measured. Diet composition estimates based on C-isotope discrimination have the major disadvantage that they cannot be taken to species level. By contrast, microhistological methods can identify many plant species in extrusa, digesta or faeces, but often a large proportion of plant fragments remains unidentifiable. Plant-wax hydrocarbons show great promise as markers for estimating diet composition and intake. However, we suggest that to be applicable in complex plant communities there is a need with this method either to recruit a wider range of wax markers (e.g. alcohols, sterols, fatty acids) or to use it in combination with other methods. We suggest that, in turn, this generates an urgent need for research on statistical aspects of the combined use of markers or methods, in relation to the error structures of the data or methods being combined and the standard errors of the resultant estimates of diet composition and intake. We conclude by discussing the extension of intake and/or diet composition measurements to the measurement of nutrient transactions within the gut, particularly in relation to the supply of absorbable nutrients.

Generic relationship between calcium intake and radiostrontium transfer to the milk of dairy ruminants.

The hypothesis is tested that there is a generic relationship between the calcium intake and the transfer of radiostrontium to milk which can be used for all dairy ruminants. In addition to the daily calcium intake, the relationship also requires values for the strontium to calcium observed ratio, which describes the discrimination in transfer of the two elements to milk (a value of 0.11 is used), and the calcium concentration in milk. The relationship had previously only been validated for dairy cattle as there were insufficient data for other ruminant species. Here, we present recently available data for dairy goats, and also a limited amount of data for sheep derived from the literature. From the comparison between these data and predicted values, we conclude that it is possible to derive a generic model of the transfer of radiostrontium to the milk of dairy ruminants.

A metabolic approach to simulating the dynamics of C-14, H-3 and S-35 in sheep tissues.

The results of a study in which groups of sheep were given single oral administrations of 14C, 3H and 35S and then slaughtered over a period of 1 year are reported. The experimental data were used to investigate the potential of metabolically based models for describing the transfer of the three radionuclides to sheep tissues. The structure of these models is based upon a simplified understanding of the transfer of the macro-elements C, H and S by processes such as respiration and protein synthesis/degradation. A consequence of this approach is that the three models have many common parameters. The models reproduced the general trends of the observations, accounting for 74%, 66%, and 58% of the observed variation in the 14C, 3H and 35S data, respectively, suggesting that they may provide a useful alternative approach to modelling the transfer of these radionuclides. The models presented are limited to the particular experimental situation for which they were developed, and further experimental work would be required to extend them. However, such metabolically based models have great potential: for example, they should be able to account for the influence of dietary intake, physiological status or the form of the radionuclide in the animals diet (e.g. tritiated water or organically bound tritium).

A model of radiostrontium transfer in dairy goats based on calcium metabolism.

A model for the transfer of radiostrontium (90Sr) in dairy goats is presented. The novel feature of the model is that it is based on the current understanding of Ca metabolism and assumes that the transfer of radiostrontium is driven by Ca transfer. Previously published models of radiostrontium transfer in animals have ignored the influence of Ca metabolism. Unknown model parameters were obtained by fitting the model to data from a study of radiostrontium and Ca transfer in goats. The model accounted for 95 and 97% of the observed variation in the data for Ca (n = 22) and radiostrontium (n = 43), respectively. Unlike previously reported models, the model presented here could be applied to simulate the effect of additional dietary Ca as a countermeasure to the contamination of milk by radiostrontium under different circumstances.

Variation in the metabolism of radiocaesium between individual sheep.

Considerable variability has been recorded in the radiocaesium activity concentration of muscle between individual sheep in the same flocks in upland areas that received fallout from the Chernobyl accident. In a previous paper we demonstrated that there is a propensity for certain sheep within a flock to be always amongst the most contaminated and others to be consistently the least contaminated. Here we report a study to determine the extent to which variation in the metabolism of radiocaesium by individual sheep may contribute to the observed variability within sheep flocks. The transfer coefficient and biological half-life of orally administered ionic radiocaesium in muscle were determined under controlled conditions in 22 ewes from an upland farm in an area of the UK which received comparatively high levels of Chernobyl fallout. There was considerable variation between individuals in both the transfer coefficient (0.19-0.56 day x kg(-1); mean 0.34 day x kg(-1)) and biological half-life in muscle (5.2-18.7 days; mean 9.8 days). Changes in liveweight during the study and feed intake together accounted for 72% of the variation in the derived transfer coefficients; liveweight change also accounted for 56% of the observed variation in biological half-life. In a subsequent study, the true absorption coefficient of radiocaesium was determined in 12 of the ewes. There was a positive correlation between transfer and true absorption coefficients (R = 0.57). We conclude that differences in the metabolism of radiocaesium will contribute to the observed variability in radiocaesium activity concentrations within sheep flocks in areas which were contaminated by Chernobyl fallout. We also suggest that for growing animals, the influence of liveweight change and feed intake on radiocaesium transfer may be greater than observed here. Similarly, in dairy cattle, for which feed intake changes considerably during the course of a lactation, large temporal variation in radiocaesium transfer to milk could be expected.

The use of dietary calcium intake of dairy ruminants to predict the transfer coefficient of radiostrontium to milk.

Transfer coefficients (the equilibrium ratio between radionuclide activity concentration in milk or meat and the daily intake of radionuclide) are widely used to predict the contamination of animal products following the release of radionuclides into the environment. For a transfer coefficient to be generally applicable, its value must be constant for a range of circumstances. However, this is not the case for radiostrontium, the behaviour of which is strongly influenced by that of the homeostatically controlled nutrient, calcium. In this study, a relationship is derived between radiostrontium transfer coefficients and dietary calcium intake which takes into account the observed ratio for strontium:calcium transfer to milk. This relationship is tested against a range of observed data collated from the literature (n = 30) and found to account for 93% of the variability in transfer coefficient values. Model calculations show that a reduction in Fm of at least 40%-60% would be expected if dairy cattle, fed rations typical for well-managed herds, were supplemented with 100-200 g per day. Larger reductions would be predicted when dietary calcium intake is low.

Use of the true absorption coefficient as a measure of bioavailability of radiocaesium in ruminants.

Limitations of existing methods to describe the bioavailability of dietary radionuclides to ruminants (the transfer coefficient and apparent absorption coefficient) have led to the alternative suggestion of using the true absorption coefficient (A(t)). Various approaches to estimating A(t) for radiocaesium, involving the intravenous administration of a second isotope, are presented and discussed with reference to results from studies in which a range of radiocaesium sources were examined in sheep. Although estimates of A(t) differed between the sources, they were reasonably consistent between measurement techniques. Those methods which involved the estimation of endogenous faecal excretion of radiocaesium could be used with previously contaminated animals and did not require continuous administrations of radiocaesium isotopes, but gave unreliable results for sources of low bioavailability. Methods based on estimating the turnover rate of dietary radiocaesium through blood plasma were sufficiently sensitive to measure A(t) for the range of sources studied. However, they require previously uncontaminated animals and continuous administration of both isotopes for approximately 7 days. Bioavailability is more effectively measured as A(t) than as the transfer or apparent absorption coefficients since A(t) does not incorporate factors relating to the metabolism of radiocaesium in the tissues of the animal. The results of these studies show that differences in transfer coefficients between sheep and cattle and between sheep of differing ages are not due to variation in absorption across the gut. The potential for applying these approaches to other radioactive elements is discussed.