Effects of lactic acid bacteria in inoculants on changes in amino acid composition during ensilage of sterile and non-sterile ryegrass.

A study was carried out on the changes occurring in the amino acid fraction of a hybrid ryegrass during ensilage in laboratory-scale silos to help to establish the relative roles of plant and microbial proteases on protein degradation in the silo. Herbage treatments included (i) normal grass without treatment (ii) lambda-irradiated grass (sterile) without treatment (iii) sterile, inoculated with a strain of Lactobacillus plantarum and (iv) sterile, inoculated with a strain of Lactobacillus paracasei subsp. paracasei. These treatments had a significant effect on silage amino acid profiles. Concentrations of free amino acids and the extent of amino acid catabolism varied with treatment. However, levels were notably higher in control silages after 90 days (free amino acid nitrogen constituting 54% of total amino acid nitrogen compared with 37, 32 and 22% for treatments i, ii and iv, respectively). These results indicate that the extent of protein hydrolysis during ensilage is influenced by factors other than rate of pH decline and plant protease activity, and that microbial proteases play a role.

Problems in the clinical use of intrapartum fetal ECG monitoring.

Commercial machines are now available to monitor the fetal electrocardiograph in labour (Cinventa, Sweden). We report our experience of the first one hundred women we monitored by this method. They were divided into five groups dictated by the change in the fetal scalp electrode used for monitoring and the changes made in the computer software used for signal processing. There were progressive significant improvements in the ability of the system to produce a continuous heart rate trace such that it became acceptable for routine fetal monitoring. The quality of the electrocardiogram improved to 50% of the 'check ECG complexes' being printed accurately. Electrical signal distortion causing baseline wander is however the significant remaining problem. On-line analysis of the T/QRS ratio improved very significantly from 36% to 84% of the monitoring time. There were no significant improvements in the recordings made in the second stage, which remained poorer in quality and reliability in all groups. As well as changes in the computer software, the use of a single helix electrode and practical experience contributed to the improvements.

The influence of insulin and amino acid supply on amino acid uptake by the lactating bovine mammary gland.

Four lactating dairy cows received arterial infusions of insulin (1.41 U/h), an AA mixture (threonine, methionine, leucine, phenylalanine, and lysine at 5.87, 1.90, 3.55, 2.17, and 4.21 mmol/h, respectively), and a combination of the two in a 4 x 4 Latin square. The infusions were performed over a 3-d period directly into the extra pudic artery on both sides of the mammary gland, and samples were taken simultaneously of the downstream extra pudic arterial blood and also of subcutaneous abdominal venous blood. Blood flow was measured by dye dilution using p-amino-hippuric acid and was increased by 37% by infusion of insulin plus AA (P less than .05). Infusions of AA tended to increase the arteriovenous difference and uptake of the infused AA (P less than .05 for phenylalanine) and had varying effects on the uninfused AA. Inclusion of insulin in the AA infusion tended to increase uptake of infused AA, whereas infusions of insulin alone tended to decrease uptake. There were no significant effects of infusion on milk yield or composition.

The degradation of different protein supplements in the rumen of steers and the effects of these supplements on carbohydrate digestion.

1. Four steers with simple rumen and abomasal cannulas were given diets consisting of ground and pelleted alkali-treated straw, rolled barley and tapioca supplemented with urea (diet U) or containing single-cell protein (diet SCP), maize-gluten meal (diet MGM) or rapeseed meal (diet RSM) in place of some of the tapioca. The isoenergetic diets were given in a 4 x 4 Latin square design in eight feeds/d at 3-h intervals and provided sufficient metabolizable energy to support a growth rate of approximately 0.5 kg/d. Chromic oxide and polyethylene glycol were given as markers and appropriate samples taken from the rumen and abomasum. Flows (g/d) at the abomasum of organic matter and nitrogenous and carbohydrate constituents were calculated. 2. Rumen ammonia levels were similar with all three protein supplements at about 9 mmol/l, which was significantly lower (P less than 0.05) than that in animals on diet U (16 mmol/l). Rumen liquid outflow rates (/h) were 0.099, 0.139, 0.125 and 0.160 for diets U, SCP, MGM and RSM respectively; the difference between diet U and diet RSM was significant (P less than 0.05). Corresponding values for Cr2O3 outflow rates were 0.027, 0.032, 0.027 and 0.030/h respectively, which did not differ significantly from each other. 3. RNA, 35S and diaminopimelic acid (DAP) were used as microbial markers. Efficiencies of microbial-N (MN) synthesis, expressed as g MN/kg apparently digestible organic matter, truly digestible organic matter or carbohydrate fermented, were generally not significantly affected by the diet and averaged 29, 22 and 29 respectively based on mean RNA and 35S markers. Corresponding values derived from DAP of 22, 16 and 21 g MN/kg respectively were all significantly (P less than 0.001) lower. Using 35S as microbial marker, MN flows at the abomasum as a proportion of non-ammonia-nitrogen flow were 0.78, 0.64, 0.51 and 0.78 for diets U, SCP, MGM and RSM respectively. Derived true rumen degradability values (g/g intake) of the total dietary N were 0.91, 0.79, 0.69 and 0.90 for diets U, SCP, MGM and RSM respectively. Protein supplement degradabilities for single-cell protein, maize-gluten meal and rapeseed meal were 0.73, 0.51 and 0.98 respectively. 4. Mouth-to-abomasum digestibility coefficients of the main neutral-sugar components of dietary polysaccharides were 0.68, 0.63 and 0.61 for arabinose, xylose and cellulose-glucose on diet U.(ABSTRACT TRUNCATED AT 400 WORDS)

The effects of different sources of nitrogen supplementation on the post ruminal flows of organic matter and different nitrogenous constituents in steers.

Friesian steers, virtually protozoa free, were equipped with simple rumen and abomasal cannulas. They were given diets consisting of approximately equal proportions of ground, pelleted alkali treated straw and a rolled barley, tapioca mixture supplemented with urea + casein (UC), soybean meal (SBM), 'normal' white fishmeal (NDF) or white fishmeal designated as being of 'low' rumen degradability (LDF). The diets were isoenergetic (the protein sources replacing part of the tapioca) and they were given in amounts to supply sufficient metabolizable energy (ME) to support an average growth rate of 0.5 kg/d. Rumen degradable nitrogen (RDN): ME values were estimated to be 2.08, 1.40, 1.90 and 1.66 for diets UC, SBM, NDF and LDF respectively. RNA, alpha-epsilon-diaminopimelic acid and 35S (added as sulphate) were used as bacterial markers. Chromic oxide and polyethylene glycol (PEG) were given as flow markers and flows (g/24 h) at the abomasum of organic matter (OM) and nitrogenous constituents were calculated. Rumen volumes and ruminal liquid fractional outflow rates were measured using PEG. Samples of mixed rumen bacteria separated from strained rumen digesta from animals receiving diet UC contained significantly less DAP-N (0.322 g/kg DM) than those from animals receiving diets SBM, NDF or LDF (0.530 g/kg DM). Mean rumen volume (approximately 15 l) and liquid fractional outflow rates (approximately 0.105/h) were similar on all diets but there was appreciable variation between animals. The proportion of OM intake digested in the rumen was similar on all diets. The proportional contribution of bacterial-N to the total non-ammonia-N passing the abomasum based on mean values derived from DAP and 35S as markers was 0.57, 0.47, 0.39 and 0.31 for diets UC, SBM, NDF and LDF respectively. Corresponding values based on RNA were 0.71, 0.50, 0.48 and 0.35 respectively. Bacterial-N (RNA) flows at the abomasum were 31, 25, 26 and 20 g/d for diets UC, SBM, NDF and LDF respectively. Corresponding values for 35S and DAP were 26, 24, 21 and 18 g/d respectively. Values derived from RNA flows were consistently and significantly higher (P less than 0.01) than those based on DAP or 35S. Mean estimated efficiencies of bacterial protein synthesis (g bacterial-N/kg OM truly digested) were 15, 15, 14 and 12 for diets UC, SBM, NDF and LDF respectively.(ABSTRACT TRUNCATED AT 400 WORDS)

The simultaneous estimation of the amounts of protozoal, bacterial and dietary nitrogen entering the duodenum of steers.

Four steers were given straw and tapioca diets, twice daily, in a 4 X 4 Latin-square design. These diets, containing 4.2 g nitrogen/kg dry matter (DM), were further supplemented with either urea, decorticated groundnut meal (DCGM), untreated (UT) casein or formaldehyde-treated (FT) casein to give a total of 19.7 g N/kg DM and 10.5 MJ/kg DM daily. Concurrent samples of rumen bacteria and protozoa and abomasal digesta were collected for each period of the experiment and the concentrations of 2-aminoethyl phosphonic acid (AEPA), diaminopimelic acid (DAPA), total nitrogen (TN), total phosphorus (TP), amino acids and hexosamines were determined in the dried preparations. The nature of the dietary supplements had little effect on the concentrations of most of these constituents or on the total protozoal numbers. Abomasal digesta samples marked with polyethylene glycol (PEG) and chromic oxide for flow estimation were collected over 24 h, and the proportions of protozoal-N, bacterial-N and microbial-N estimated simultaneously using the markers AEPA, DAPA and RNA respectively. These digesta-N components were also estimated using an amino acid profiling (AAP) method which gave, in addition, estimates of the dietary and endogenous components. For the diets containing casein, the proportion of dietary casein was estimated directly using casein-P as a marker. Estimates of the respective mean proportions of microbial-N in abomasal digesta non-ammonia-N (NAN) for the diets containing urea, DCGM, UT casein or FT casein were: AEPA 0.56, 0.32, 0.27 and 0.16; DAPA 0.88, 0.70, 0.81 and 0.57; RNA 0.98, 0.85, 0.92 and 0.53. Giving FT casein significantly (P less than 0.001) increased the flow of casein-N at the abomasum and a significantly (P less than 0.001) greater proportion of casein-N was found in abomasal NAN (0.51 v. 0.09) where FT rather than UT casein was given. The AAP method gave results for the proportions of microbial- and dietary-N (where casein was given) which were, in general, slightly lower than those obtained using RNA and casein-P as markers. Agreement with estimates of bacterial protein (from DAPA) and of protozoal protein (from AEPA) was less satisfactory. Comparisons of the various estimates of the proportions of microbial-N in abomasal digesta suggested that the results obtained for protozoal-N by AEPA were overestimates. AEPA was found in mixed rumen bacteria which may have accounted in part for these overestimates. However, AEPA was not detected in any of the dietary ingredients.