Methods for the primary culture of muscle cells from fetal sheep were developed which gave high yields of cells. Myoblasts were grown in vitro, and allowed to fuse to form contractile multinucleate myotubes; these could be maintained in a good condition for at least 2 weeks. Protein turnover in these differentiated cultures was examined for sensitivity to each of four potentially anabolic peptide hormones and growth factors: insulin, insulin-like growth factor I (somatomedin C), epidermal growth factor and growth hormone. Insulin was found to have no effect except at high concentrations (1 mumol/l), compatible with its role as a somatomedin analogue. Insulin-like growth factor I was active at lower levels (1 nmol/l) but the cultures were not as responsive to it as were primary rat muscle cultures or differentiated L6 cells, which were tested in similar experiments. The maximum stimulation of protein synthesis observed with the ruminant system was only 16%. Epidermal growth factor was highly anabolic for primary cultures from sheep muscle, and the cells were very sensitive to it, half-maximal stimulation of protein synthesis being seen with concentrations as low as 20 pmol/l. No effects of bovine growth hormone were seen in the ovine system. However, an inhibition of protein breakdown was found with high concentrations (0.1 mumol/l) in the L6 rat myoblast cell line. It was found that the culture conditions used could affect the observed responses of protein synthesis and degradation, despite withdrawal of serum from the incubation media 22 h before testing.

1. The effects of Revalor (trenbolone acetate plus oestradiol) implantation or the inclusion of clenbuterol (a 8-2-adrenergic agonist) in the diet of wether lambs was studied. Using continuous intravenous infusion of [3H]tyrosine the fractional synthetic rate of mixed protein from three separate muscles was measured.2. Clenbuterol slightly increased growth rate but had a significant (P < 0.02) effect on food conversion efficiency.The weight and protein content of the longissimus dorsi and vastus lateralis muscles were increased but no such changes were observed for the vastus intermedius. For the longissirnus dorsi at least the increase was probably achieved by a reduction in fractional degradation rate of the muscle protein.3. Revalor significantly increased the growth rate and food conversion efficiency of the animals. This increase was not specific for muscle. Estimated degradation rates of muscle protein were lower in the treated animals. 4.The possible mode of action of these materials was discussed. The results obtained again highlight the importance of protein degradation in controlling growth.

The effect of supplementing grass silage with fishmeal on growth, muscle composition and the rate of muscle protein synthesis was investigated in young Friesian steers with and without oestradiol implants. The effect of the P-adrenergic agonist cimaterol was simultaneously investigated in animals fed on silage alone. Treatments lasted for 9 or 10 weeks. Fishmeal supplementation significantly increased animal growth rates (P < 0.001) and the weights of three dissected muscles (P < 0.001) compared with the silage-fed controls. These effects were further enhanced in animals also implanted with oestradiol. Muscle weights expressed as a proportion of body-weight were increased by fishmeal, suggesting that protein deposition had been enhanced. No further increase in the proportional muscle weights was obtained with oestradiol. Muscle dry matter content tended to be increased in both implanted and nonimplanted animals receiving fishmeal compared with controls, but the proportions of protein, fat and ash were relatively constant. The intramuscular lipid composition was slightly altered by fishmeal. Muscle protein fractional synthetic rates (FSR), measured by continuous infusion of [3H]tyrosine, were increased by fishmeal in all three muscles of both implanted and non-implanted animals. There were no differences, however, due to oestradiol, over non-implanted fishmeal animals. This suggests that oestradiol may increase muscle accretion by reducing protein degradation rate. Cimaterol significantly increased longissimus dorsi (P < 0.05) and vastus lateralis (P < 0.01) muscle weights but had no effect on semitendinosus muscle weight or live-weight gain. The proportion of protein was increased (P < 0.001) and the fat content reduced (P < 0.05) in all three muscles but intramuscular lipid composition was not markedly affected. Whilst methylhistidine : creatinine excretion was reduced by cimaterol, FSR were increased in the 1. dorsi and v. lateralis muscles suggesting P-agonists have effects on both protein synthesis and protein degradation.

Three body amino acid pools (plasma free, plasma bound and small intestinal tissue) were evaluated as precursors to allow measurement using the isotope dilution technique of endogenous excretion at the terminal ileum of animals. Eighteen 150-g bodyweight rats were given either a protein-free, an enzyme-hydrolysed casein based or a synthetic amino acid based diet, and digesta were collected from the terminal ileum. The animals had been subjected to a constant 8-day infusion of tritiated leucine via subcutaneously implanted osmotic mini pumps. Specific activities (dpm nM-' leucine) of the ileal digesta and the plasma free, plasma bound and small intestinal tissue pools were determined and the specific activity for the ileal digesta was expressed as a proportion of the respective precursor pool value to give dilution factors for each dietary treatment. For the protein-free diet, where the ileal nitrogenous flow is endogenous by definition, the dilution factor for an appropriate precursor pool would be unity.For the hydrolysed casein and synthetic amino acid diets, in which the peptides and amino acids are expected to be virtually completely absorbed anterior to the ileum, high dilution factors (close to unity) would be expected. The mean dilution factors based on the plasma free amino acid pool were untenably low (0.2 to 0.3). For the plasma bound amino acid pool mean dilution factors of 1.3 were found for animals given the protein-free and synthetic amino acid diets, while a lower value (0.7) was obtained for the hydrolysed casein treatment. Untenably high factors (1.5) were found with the small intestinal tissue for the protein-free and synthetic amino acid treatments, while the corresponding value for the hydrolysed caseinfed rats was unity. The dilution factor data within treatments were highly variable, and none of the pools examined gave consistently reliable results and could thus be accepted as a valid precursor pool for the endogenous proteins.

SU MMARYBy a variety of exposure routes it is possible that the toxic heavy metals cadmium, arsenic and mercury could enter the diet of farm animals and hence contaminate food products derived from those animals. Therefore, there is a need to be able to assess the likely levels of contamination in animal tissues if exposed to contaminated feed and also to estimate how rapidly an animal will decontaminate once the source of contamination is removed from the feed. The development of dynamic models to predict changes in the degree of heavy metal contamination in tissues of ruminants have been hindered by the lack of data on the transfer and excretion rates of these metals from tissues. A study is described during which dairy cows were given a single intraruminal administration of 109 Cd, 73 As and 203 Hg and measurements made of the subsequent concentrations of the radioisotopes in body tissues and milk. The resultant data were used to adapt previously developed compartment models describing the behaviour of the metals in sheep for use with dairy cows. Two changes were made to the sheep models : (i) a new excretion route was included to describe transfer to milk; (ii) the rate coefficients (with the exception of those involving gut absorption and transfer) were adjusted according to the ratio of the metabolic live-weights (live-weight (kg) 0 . 75 ) of the sheep to that of the cattle. The models predicted levels of the metals in the cattle tissues and milk reasonably well.