Role of rumen protozoa in nitrogen digestion in sheep given two isonitrogenous diets

1. The effect of protozoa on digestion in the rumen was studied using either defaunated or faunated sheep. 2. Six wethers, each fitted with rumen and simple duodenal cannulas, were given two isonitrogenous diets containing either lucerne (Medicago sativa) hay (diet L) or sodium hydroxide-treated wheat straw (diet S). The diets were given in eight equal portions per day at 3-h intervals. The mean intake of dry matter, 53 g/kg body-weight0.75 per d, was similar for the two diets and each diet had a similar digestible organic matter content. Diet L promoted a large protozoal population and was rich in nitrogen sources of low rumen-degradability, while diet S supported a smaller protozoal population and was rich in rumen-degradable N. 3. Digesta flow at the duodenum was estimated by means of a dual-marker technique using chromium-mordanted lucerne hay and polyethylene glycol as markers. The microbial flow at the duodenum was estimated using diaminopimelic acid (DAPA), nucleic-acid purine bases (PB) and 35S incorporation simultaneously. The different microbial markers were compared in the defaunated sheep. Protozoal N contribution was estimated in faunated sheep. 4. Defaunated sheep had lower rumen ammonia concentrations and molar proportions of butyric acid than faunated sheep, but they had higher molar proportions of propionic acid. 5. Rumen organic matter digestion was reduced by defaunation, but this decrease was compensated for by increased intestinal digestion. 6. There was a net increase of N flow (approximately 10 g/d) between mouth and duodenum in defaunated sheep. This was explained by increases in both microbial and dietary N flows from the rumen compared with faunated sheep. 7. The influence of protozoa on solid- and liquid-phase retention times in the rumen is discussed, as well as the protozoal contribution to microbial N flow in the duodenum of faunated sheep.     

Dynamics of protozoa in the rumen of sheep

Protozoa were labelled by incubating 100 ml rumen fluid with [14C]choline for 1 h. The protozoa were concentrated by centrifugation and then washed with rumen fluid. This reduced residual 14C in the fluid medium to insignificant amounts while still retaining the viability of the labelled protozoa. Washing procedures using formal saline (40 g formaldehyde/1 saline (9 g sodium chloride/1)) and saline were developed to isolate protozoa for estimation of specific radioactivity. 2. The protozoal pool in freshly-collected rumen fluid incubated in vitro retained 90% of the radioactivity for up to 6 h following addition of 14C-labelled protozoa produced as indicated previously. The specific radioactivity of protozoa did not change during the incubation period. 3. Protozoa labelled with [14C]choline and then stored until they died rapidly lost 14C to methane when they were incubated in rumen fluid or were injected into the rumen. Some [14C]choline was salvaged under these conditions by the live protozoa present as they apparently incorporated up to 13% of the label from the dead protozoa. However, protozoal debris from the injected solution could also have been present in the isolated protozoa. 4. The in vitro results suggested that the protozoal preparations were viable, and that the incorporated choline did not have a turnover in excess of the turnover of nitrogen (i.e. specific radioactivity remained constant with time in vitro) suggesting that the dilution of specific radioactivity of protozoa following mixing of a 14C-labelled dose of protozoa represented the rate of irreversible loss and also replacement of protozoa in the rumen. 5. 14C-labelled protozoa had a half-life in the rumen which was greater than that of rumen fluid and in six animals the protozoal replacement rate was 1-4.1 mg N/min. 6. Losses of 14C from labelled protozoa in the rumen in methane or via abomasal digesta were 65 and 35% respectively. 7. The results suggest that protozoal growth may be as high as 32% of the total microbial protein synthesis in the rumen but that 65% of the protozoa die and are degraded in the rumen.     

Dynamics of protozoa in the rumen of cattle

1. The dynamics of protozoa were studied in two groups of rumen-fistulated cattle fed on a basal diet of molasses ad lib., with oaten chaff given at 6 or 18 g/kg live weight. This diet resulted in different mixtures of protozoal species in the populations in the rumen. 2. The rumen protozoa were studied by intrarumen injections of protozoa labelled in vitro with [14CH3]choline. An indication of protozoal death and fermentation of protozoal cell residues was obtained by measuring 14C loss via the methane pool. 3. After a single injection of labelled protozoa, the decline in the specific radioactivity (microCi/g nitrogen) of the protozoal pool in the rumen indicated that first-order kinetic processes applied. Conversely the specific radioactivity of protozoa, incubated in rumen fluid, remained constant indicating no growth in vitro, presumably owing to a rapid exhaustion of essential nutrients. 4. The protozoal populations in the rumen of cattle fed on the diet with the low level of oaten chaff were mainly small ciliates; but on the higher level of chaff in the diet, the large ciliates were a higher proportion of the total protozoal population present. 5. The mean pool size of protozoa in the rumen was significantly larger and the protozoal half-life tended to be longer for cattle fed on the higher level of chaff in the diet. The apparent production rate of protozoa in cattle fed on each diet was not significantly different and there were no differences in the production rate of methane. The percentage losses of label from protozoa in the rumen via the methane pool were not significantly different on the two diets and indicated that 74% of the protozoa that were apparently irreversibly lost from the rumen could be accounted for by death and lysis in the rumen and therefore only 26% of protozoa apparently entered the lower digestive tract.     

The relationships between potassium intakes, transmural potential difference of the rumen epithelium and magnesium absorption in wethers

In vitro studies with isolated sheep rumen epithelium have shown that an increase in the lumen K concentration induces an increase in the transmural potential difference across the rumen epithelium (serosal side: positive), which is associated with a decrease in Mg transport. However, at lumen K concentrations >80 mmol/l, Mg transport across the epithelium became independent of the lumen K concentration. The present study was carried out to determine whether this observation also occurs in vivo. Four ruminally fistulated wethers were fed four rations supplemented with KHCO3 (15.7, 37.6, 59.4 or 77.4 g K/kg DM) in a 4 x 4 Latin square design. Increased K intakes significantly increased the rumen K concentration. For all data combined, Mg absorption expressed as % intake was negatively correlated with the rumen K concentration. However, apparent Mg absorption either expressed in absolute terms (g/d) or as % intake was not significantly affected when the dietary K concentration was increased from 59.4 to 77.4 g/kg DM. Rumen K concentration was inversely correlated with the transmural potential difference (blood side: positive) (Pearson's r -0.709; R(2)adj 0.468, P=0.002, n 16). It is concluded that in wethers apparent Mg absorption becomes independent of the dietary K concentration when the K concentration is >60 g/kg DM or equivalent to a postprandial rumen K concentration of about 125 mmol/l.     

Dynamics of large ciliate protozoa in the rumen of cattle fed on diets of freshly cut grass

1. The dynamics of large ciliate (holotrich) protozoa (Isotricha and Dasytricha spp.) in the rumen of cattle given cut, fresh ryegrass (Lolium multiflorium Lam) were studied by means of a single intrarumen injection of 14C-labelled protozoa prepared in vitro by adding [Me14C]choline to rumen fluid containing protozoa and incubating at 39 degrees for 2 h. 2. An indication of the lysis rate of protozoa in the rumen was obtained from the radioactivity apparently lost through the methane pool. 3. The turnover time of the holotrich protozoa indicates that these protozoa were extensively retained in the rumen and that only a small proportion of those produced in the rumen flowed out in the digesta. This was supported by the estimation of the rate of lysis which was approximately 85% of the turnover rate in the rumen. 4. The apparent production rate of the larger protozoa indicates that they contribute only about 9% of the predicted net microbial protein synthesis in the rumen.     

The contribution of protozoa to the protein entering the duodenum of sheep.

1. Four sheep, each fitted with a rumen fistula and a re-entrant cannula at the proximal duodenum were fed a semi-purified diet containing urea as the only nitrogen source. The quantities of total protozoal amino acid-N (TPAN) present in the rumen and entering the duodenum were determined when the mean rumen dilution rate (D) was low (0.034/h) and when D was increased to 0.078/h by the intraruminal infusion of artificial saliva. 2. Increasing the dilution rate had no significant effect upon the proportions of TPAN present in the total microbial amino-acid-N (TMAN) of the rumen fluid and duodenal digesta. With both dilution rates the mean proportion of TPAN in the duodenal TMAN (0.24) was markedly less than the equivalent proportion (0.45) found in the rumen fluid. 3. The daily flow of TPAN, as measured at the duodenal cannula at both dilution rates were equivalent to only 41% of the flow of TPAN as predicted from measurements of rumen outflow, indicating that a substantial proportion of rumen protozoal protein was retained within the rumen.     

Changes in the rumen metabolism of sheep given increasing amounts of linseed oil in their diet.

1. Linseed oil was incorporated gradually into the diet of four sheep until the animals received 90 g additional fat/d. Attempts were made to measure changes in concentration of substances and rates of synthesis in the rumen directly, and by incubation of rumen contents in vitro (zero-time technique). 2. The high-fat diet increased the dilution rate and the volume of rumen contents and decreased the synthesis of diaminopimelic acid in the rumen. The number of protozoa decreased and the number of bacteria increased in the rumen of animals receiving the high-fat diet. 3. The concentration of volatile fatty acids (VFA) in the rumen decreased for sheep given the high-fat diet, but the capacity of rumen contents to produce VFA in vitro increased. 4. The incorporation of radioactivity from [14C]acetate into lipids during incubation of rumen contents vitro increased with the amount of linseed oil in the diet. The greatest proportional increase was with the bacterial fraction of rumen contents. 5. In the group of four animals used, one animal showed consistent differences in the magnitude of the measured varibles. This animal appeared to have a smaller rumen, a lower dilution rate and larger concentrations of some substances in the rumen. A higher proportion of fatty acids appeared to be synthsized by the micro-organisms from this animal.     

Reponses of sheep to a vaccination of entodinial or mixed rumen protozoal antigens to reduce rumen protozoal numbers

Two rumen protozoa vaccine formulations containing either whole fixed Entodinium or mixed rumen protozoa cells were tested on Merino sheep with the aim of decreasing the number and/or activity of protozoa in the rumen. Negative control (no antigen) and positive control (Tetrahymena corlissi antigens) treatments were also included in the experiment. Blood and saliva were sampled to measure the specific immune response. Protozoal numbers in the rumen were monitored by microscopic counts. Vaccination with protozoal formulations resulted in the presence of specific IgG in plasma and saliva, but saliva titres were low. Titres after secondary vaccination were higher (P 0.05) by the vaccination and there was also no difference (P>0.05) between treatments in rumen fluid ammonia-N concentration or wool growth. In vitro studies investigated the binding ability of the antibodies and estimated the amount of antibody required to reduce cell numbers in the rumen. The studies showed that the antibodies did bind to and reduced protozoa numbers, but the amount of antibody generated by vaccination was not enough to produce results in an in vivo system. It is suggested that the vaccine could be improved if specific protozoal antigens are determined and isolated and that improved understanding of the actions of protozoa antibodies in rumen fluid and the relationships between levels of antibodies and numbers of protozoa in the rumen is needed.     

Comparison of non-tracer and tracer methods for determination of volatile fatty acid production rate in the rumen of sheep fed on two levels of intake

The aim of the present work was to estimate volatile fatty acid (VFA) production rate in the rumen of sheep fed two levels of intake using both a tracer (TM; by isotope dilution) and a non-tracer method (NTM; by supplementary infusion) in steady-state conditions. Six wethers received a diet containing 700 g lucerne hay and 300 g ground maize/kg in eight equal meals at 3 h intervals per d. The diet (9.8 MJ metabolizable energy (ME)/kg DM) was offered at 90 % ad libitum consumption (high intake, HI) or 45 % ad libitum consumption (low intake, LI) in a crossover design. Each sheep received five intrarumen VFA solutions infused continuously for 24 h at rates of 250 ml and 165 ml/h for the HI and LI respectively. The first infusion, considered as a control treatment (Con), consisted of a solution of [1-(13)C]propionate (7 mmol/d). The four other solutions were isoenergetic (1.9 MJ ME/kg DM intake) mixtures of unlabelled propionate (C(3)) and butyrate (C(4)) at different levels: 0.90 mol C(4)/kg DM intake; 0.60 mol C(3)/kg DM intake; 0.30 mol C(3)/kg DM intake; 1.35 mol C(3)/kg DM intake. The VFA infusions did not affect rumen fermentation of the basal diet (pH, osmotic pressure, protozoa numbers), and comparable DM digestibility of the diet among the different treatments was observed. Both estimation methods demonstrated a similar increase (1.7-fold) in the rumen VFA production rate of sheep fed at intakes varying between 0.9 to 1.7 times maintenance. Irrespective of the intake level, the rumen production rate of individual VFA was on average 1.5-fold higher when estimated by the TM compared with the NTM. Rumen VFA production rates estimated by the NTM and TM represented 80 % and 120 % ME intake respectively. The difference between NTM and TM estimates seems likely to be caused mainly by overestimation of the VFA production rates by the TM.     

Contribution of rumen protozoa to duodenal flow of nitrogen, conjugated linoleic acid and vaccenic acid in steers fed silages differing in their water-soluble carbohydrate content

The present experiment was designed to estimate the quantitative contribution of rumen protozoa to the total N, conjugated linoleic acid (CLA) and vaccenic acid (VA; trans-11-18 : 1) flow to the duodenum of steers fed two silage diets: control silage (CS) and silage high in water-soluble carbohydrates (HS). Protozoal duodenal flows were estimated using a real-time PCR assay to quantify the genes encoding protozoal 18S ribosomal RNA. Denaturing gradient gel electrophoresis was used to confirm that the rumen protozoa populations were similar to the protozoal population flowing to the duodenum. Estimated duodenal flow of protozoal N was 14.2 and 18.2 g/d (P>0.05) for animals fed the CS and HS diets respectively. Protozoal flow thus represented between 12 and 15 % of the total N duodenal flow. In terms of fatty acid flow, protozoa accounted for between 30 and 43 % of the CLA and 40 % of the VA reaching the duodenum. The contribution of protozoa to 16 : 0 and 18 : 0 flows to the duodenum was less than 20 and 10 %, respectively. These results show that the fatty acids within protozoa make up a significant proportion of the CLA and VA reaching the duodenum of ruminants.     

The effect of varying the amount of linseed oil supplementation on rumen metabolism in sheep

The effects of three levels of linseed oil (LSO) supplementation of a basal diet on rumen digestion and flow of nutrients to the proximal duodenum of three mature sheep provided with permanent rumen and duodenal re-entrant cannulas were studied. 2. A basal diet of 200 g hay and 400 g concentrates daily, providing approximately 7.0 MJ digestible energy and 13 g N/d, was given alone or with supplements of 13, 26, or 40 ml LSO/d in two equal portions of 06.00 and 18.00 hours. The flow of duodenal digesta was measured by spot-sampling using chromic oxide paper as the marker. Bacterial protein synthesis (BPS) was measured by the diaminopimelic acid technique. 3. Addition of LSO reduced the digestion of energy and organic matter, particularly acid-detergent fibre, in the stomach. Digestion in the intestines increased but at the higher levels of supplementation this failed to compensate completely for the reduction in rumen digestion. Total volatile fatty acid concentrations were not affected but molar proportions of acetate and butyrate were decreased by approximately 18 and 61% respectively while the molar proportion of propionate was increased twofold by the highest concentration of oil. The higher concentrations of LSO virtually eliminated protozoa from the rumen. 4. The second increment of LSO (26 ml/d) produced the highest duodenal flow of total N and bacterial N and the highest efficiency of BPS. The highest concentration of oil (40 ml/d) was without effect. Rumen and duodenal ammonia concentrations and plasma urea concentrations tended to be reduced by the higher concentrations of LSO. 5. It is argued that the results support suggestions made elsewhere that free oils reduce the efficiency of BPS but that they also reduce the numbers of protozoa which can cause an increase in efficiency of BPS. The net effect of free oil supplementation on BPS is thus likely to be variable and difficult to predict.     

Rumen ciliate protozoa contain high concentrations of conjugated linoleic acids and vaccenic acid, yet do not hydrogenate linoleic acid or desaturate stearic acid

Conjugated linoleic acids (CLA) have been shown to improve human health. They are derived from the microbial conversion of dietary linoleic acid (cis-9,cis-12-18 : 2 (LA)) in the rumen. An investigation was undertaken to determine the role of ruminal ciliate protozoa v. bacteria in the formation of CLA and its precursor in animal tissues, vaccenic acid (trans-11-18 : 1 (VA)). Mixed protozoa from the sheep rumen contained at least two to three times more unsaturated fatty acids, including CLA and VA, than bacteria. Different species had different composition, with larger fibrolytic species such as Epidinium ecaudatum caudatum containing more than ten times more CLA and VA than some small species, including Entodinium nanellum. In incubations with ruminal microbial fractions (bacterial fraction (BAC), protozoal fraction (PRO)), LA metabolism was very similar in strained ruminal fluid (SRF) and in the BAC, while the PRO had LA-metabolising activity an order of magnitude lower. Using PCR-based methods, no genes homologous to fatty acid desaturase genes were found in cDNA libraries from ruminal protozoa. The absence of an alternative route of VA/CLA formation via desaturation of stearate was confirmed by incubations of SRF, BAC or PRO with [14C]stearate. Thus, although protozoa are rich in CLA and VA, they appear to lack the ability to form these two fatty acids from LA or stearate. The most likely explanation is that protozoa preferentially incorporate CLA and VA formed by bacteria. The implication of the present findings is that the flow of unsaturated fatty acids, including CLA and VA, from the rumen could depend on the flow of protozoa rather than bacteria.     

Factors affecting the rate of breakdown of bacterial protein in rumen fluid

1. The cellular proteins of Butyrivibrio fibrisolvens, Lactobacillus casei, Megasphaera elsdenii, Selenomonas ruminantium and Streptococcus bovis were labelled by growth in the presence of L-[14C]leucine, and the breakdown of labelled protein was measured in incubations of these bacteria with rumen fluid to which unlabelled 5 mM-L-leucine was added. The rate of protein breakdown was estimated from the rate of release of radioactivity into acid-soluble material. 2. Protein breakdown occurred at different rates in different species. The mean rates for B. fibrisolvens, L. casei, M. elsdenii, Sel. ruminantium and Str. bovis were 28.6, 18.1, 17.7, 10.5 and 5.3%/h respectively in samples of strained rumen fluid (SRF) with different protozoal populations. Rates of 3%/h or less were found in SRF from ciliate-free sheep or in faunated SRF from which protozoa had been removed by centrifugation. Further removal of mixed rumen bacteria had little effect. Suspensions of washed protozoa degraded bacterial protein at rates which were of the same order as those found in SRF. 3. The rate of breakdown of bacterial protein in different samples of SRF tended to increase as the numbers of small entodiniomorphid protozoa increased. The numbers of larger entodiniomorphs and holotrichs had no obvious influence on this rate. 4. Autoclaved and u.v.-treated bacteria were generally no different from live bacteria in their susceptibility to breakdown in SRF from faunated sheep, indicating that endogenous protein turnover was not a significant cause of bacterial protein catabolism. 5. The rate of bacterial protein breakdown was unrelated to the proteolytic activity of SRF. 6. It was concluded that predation by small protozoa is by far the most important cause of bacterial protein turnover in the rumen, with autolysis, other lytic factors and endogenous proteolysis being of minor importance.     

New perspectives on the use of tropical plants to improve ruminant nutrition

Inadequate nutrition is the main cause of low productivity by ruminants in sub-Saharan Africa. The primary feed resources in the region include natural pasture and crop residues that have tough texture, poor digestibility and are deficient in nutrients. These deficiencies can be corrected by supplementation with high-density feeds such as oilseed cakes and proteins of animal origin. However, protein sources such as oilseed cakes are beyond the economic reach of most farmers, while the incidence of bovine spongiform encephalopathy in Western intensive animal production may be thought to argue against the use of animal proteins. Local tree legumes have been investigated as potential supplements for ruminants because of their beneficial effect of increasing metabolizable energy intake, N intake and feed efficiency, and improving animal performance. However, our work has suggested that some plant materials may have a nutritional value beyond simply their nutrient content, i.e. as rumen-manipulating agents. The foliage of some tree legumes has been shown to be selectively toxic to rumen protozoa. Rumen protozoa ingest and digest bacteria and fungi, degrading their cellular protein to NH3. Microbial protein turnover due to protozoal predation in the rumen may result in the net microbial protein outflow being less than half the total protein synthesized. Results from in vivo experiments have clearly shown that duodenal flow of both undegraded dietary and bacterial protein is generally increased by defaunation. However, no practical method has been developed to date to eliminate protozoa. Anti-protozoal plants may be promising, safe, natural defaunating agents.     

The role of rumen protozoa in the utilization of paspalum (Paspalum dilatatum) hay by cattle

Six Friesian heifers (250 kg live weight) with permanent cannulas in the rumen and abomasum were allocated at random into two groups of three. One group was treated with Teric GN9 (ICI (Aust.) Ltd) to defaunate the animals during the first two of the four periods of the experiment, after which they were refaunated. The second group was treated with Teric at the end of the first two periods. The dietary treatments were: paspalum (Paspalum dilatatum) hay (4.1 kg/d) given alone and the hay supplemented with urea (20 g/kg dry matter). Defaunation was not complete but the approximate volume of protozoa in the rumen of treated animals was less than 6% of that in the untreated animals. The amount of organic matter (OM) digested in the stomach was lower (P less than 0.01) in animals with reduced fauna than in those with normal fauna. There were reductions in both the apparent OM digestibility in the total tract (from 0.56 to 0.52, P less than 0.01) and the proportion of the digestible OM digested in the rumen (from 0.82 to 0.79, not significant) of animals with reduced fauna. Apparent digestibilities of acid-detergent fibre and neutral-detergent fibre were significantly lower (P less than 0.01) in animals with reduced fauna. The amount of nitrogen disappearing from the stomach was significantly higher (P less than 0.01) with the urea supplement; effects due to concentrations of protozoa were not significant. The flow of non-ammonia-N from the abomasum was higher (P less than 0.05) in animals with reduced fauna than in animals with normal fauna. The flows of bacterial N from the abomasum and the efficiencies of bacterial N synthesis were not significantly affected by the treatments. N retention was higher (P less than 0.01) in animals receiving the urea supplement but differences due to protozoa were not significant. Protozoal contribution to the microbial N flowing from the rumen of animals with normal fauna was estimated to be 24 and 27% with and without the urea supplement respectively. Concentrations of rumen-fluid ammonia-N were reduced (P less than 0.05) and those of volatile fatty acids were increased (P less than 0.01) with reduction in protozoal numbers. Molar proportions of propionic acid increased (P less than 0.05) and of butyric acid decreased (P less than 0.01) with reduced rumen fauna.(ABSTRACT TRUNCATED AT 250 WORDS)     

Duodenal flow and digestibility in fauna-free sheep and in sheep monofaunated with Entodinium caudatum or Polyplastron multivesiculatum

Three groups of five rumen and duodenum cannulated fauna-free sheep were used in a 28 d experiment. One group remained fauna-free, whereas the second (EN) and third (PP) groups, respectively, were inoculated intraruminally with the protozoan species Entodinium caudatum and Polyplastron multivesiculatum. Rumen fluid, duodenal digesta and faecal samples were collected during the last 12 d. The flow of digesta to the duodenum was determined using Yb and Co as dual-phase markers. (15)Nitrogen and phosphatidylcholine were used as markers to calculate the duodenal flow of bacterial and protozoal N, respectively. Results showed an increase (P < 0.1) in the rumen concentration of NH3-N and total volatile fatty acids, and a decrease (P < 0.05) in the duodenal flow of non-NH3-N and bacterial N in sheep with EN and PP monofaunas, compared with fauna-free sheep. There were no differences (P > 0.05) in these variables between the two monofauna groups. Protozoal N accounted for 8 % of the duodenal non-NH3-N flow in the EN-monofaunated sheep, whereas no such flow was detected in the PP-monofaunated sheep. Apparent rumen digestibility of organic matter, neutral detergent fibre and acid detergent fibre were similar (P > 0.05) in the monofaunated groups of sheep, but rumen acid detergent fibre digestibility was higher (P < 0.05) in the monofaunated than in the fauna-free groups. Experimental results suggested that, unlike EN, the PP monofauna might not contribute to the duodenal flow of microbial protein, whereas both monofaunas showed a virtually equal degree of predation on rumen bacteria.     

Transgenic maize in the presence of ampicillin modifies the metabolic profile and microbial population structure of bovine rumen fluid in vitro

Recently, transgenic crops have been considered as possible donors of transgenes that could be taken up by micro-organisms under appropriate conditions. In an in vitro rumen simulation system, effects of ampicillin on microbial communities growing either on rumen contents with transgenic maize carrying a gene that confers resistance to ampicillin or its isogenic counterpart as substrates were examined continuously over 13 d. Rate of production of SCFA was measured to determine functional changes in the rumen model and single-strand conformational polymorphism was used to detect alterations in structure of the microbial community. Rumen contents treated with ampicillin displayed a marked decrease in the rate of production of SCFA and diversity of the microbial community was reduced severely. In the presence of transgenic maize, however, the patterns of change of rumen micro-organisms and their metabolic profiles were different from that of rumen fluid incorporating maize bred conventionally. Recovery of propionate production was observed both in the rumen fluid fed transgenic and conventional maize after a delay of several days but recovery occurred earlier in fermenters fed transgenic maize. Alterations in the microbial population structures resulting from the ampicillin challenge were not reversed during the experimental run although there was evidence of adaptation of the microbial communities over time in the presence of the antibiotic, showing that populations with different microbial structures could resume a pre-challenge metabolic profile following the introduction of ampicillin, irrespective of the source of the plant material in the growth medium.     

Effects of feed intake on composition of sheep rumen contents and their microbial population size

The present study was conducted to determine the effect of feed intake on the composition of the rumen contents of sheep and on their bacterial densities. Whole rumen contents were sampled after a period of continuous inter-rumen infusion of 15NH3 from four rumen-cannulated wethers successively fed on a hay-concentrate diet (2:1, w/w on a DM basis) at two rates of feed intake: 40 and 80 g DM/kg body weight0.75. Total weight and chemical composition of rumen contents, as well as the distribution by size and chemical composition of particles, were determined. The populations of bacteria associated with the liquid (liquid-associated bacteria, LAB) and solid (solid-associated bacteria, SAB) fractions of rumen digesta and the distribution of SAB according to feed particle size were also examined. The greater feed intake caused an increase in the mass of the rumen contents, while its chemical composition did not change, except for a higher content of organic matter (P=0.023). The distribution of feed particles by size was similar at both levels of intake. The concentrations of neutral- and acid-detergent fibre in feed particles decreased and those of total, dietary, and microbial N increased, both with a quadratic response (P=0.001), as particle size decreased. The proportion of LAB in the microbial biomass of rumen digesta reached only 8.0 %. This proportion and the density of LAB were unaffected by the level of feed intake, whereas an apparent reduction (10.4 %) occurred with the SAB biomass in whole rumen contents. A systematic, but not significant, reduction (mean value 11.9 %) in the level of microbial colonisation in the different particle fractions with the increase of feed intake was also observed.     

Effect of defaunation on protein and fibre digestion in sheep fed on ammonia-treated straw-based diets with or without maize

Using a defaunating method which preserved bacteria and fungi in the rumen, the effect of protozoa on protein and fibre digestion was studied in six adult wethers in relation to the nature of the diet. Sheep were given daily, 42 g dry matter (DM)/kg metabolic body-weight (W0.75), one of two isonitrogenous diets: one contained ammonia-treated wheat straw as the only energy source (diet S) and the other was supplemented with maize grain pellets (diet SM). Mean daily intakes (g/d) of nitrogen, neutral-detergent fibre and acid-detergent fibre were respectively 22, 573 and 373 for diet S and 23, 450 and 334 for diet SM. Elimination of protozoa increased duodenal non-ammonia-nitrogen flow. This result was mainly due to an increase in microbial protein flow and, to a lesser extent, to a higher dietary protein flow. Defaunation markedly increased the efficiency of microbial protein synthesis. Maize-grain supplementation had a net positive effect on this variable in defaunated sheep, but not in faunated sheep. Cell-wall carbohydrates were less well digested in the defaunated rumen, and the negative effect of defaunation was greatest with the diet SM. Intestinal fibre digestion increased in the defaunated sheep especially in those fed on diet SM, but not enough to compensate for the decrease in rumen digestion.     

Studies in sheep on the absorption of magnesium from a low molecular weight fraction of the reticulo-rumen contents

1. Six sheep, three animals per diet, were prepared with rumen fistulas and fed on frozen grass or grass-maize pellets to give magnesium intakes of 1.79 and 2.23 g/d respectively. The mean apparent availabilities of Mg in sheep fed on frozen grass and grass-maize pellets were 0.31 and 0.36 respectively. 2. The rumen contents were fractionated by straining the digesta through linen cloth and then differentially centrifuged to give 20,000 g and 100,000 g supernatant fractions. 3. In all sheep, regardless of diet, at 4 and 16 h after a meal, 50 and 60% respectively of the total Mg in the rumen contents was found in strained rumen fluid while 30 and 38% respectively of the total Mg was found in the 100,000 g supernatant fraction. 4. The net absorption of Mg from the temporarily isolated and washed reticulo-rumen was studied using either 100,000 g supernatant fractions of rumen contents from sheep fed on one or other of the two diets, or inorganic buffers containing the same concentration of Mg and other macroelements. 5. The Mg was readily absorbed from the 100,000 g supernatant fraction placed in the rumen with the rate of absorption being 7.3 mumol/l per min (505 mg/d) from the supernatant fraction obtained from sheep fed on frozen grass and 11.3 mumol/l per min (781 mg/d) from the supernatant fraction from sheep fed on grass-maize pellets. In the same sheep, the previously described rates of Mg absorption from the 100,000 g supernatant fraction were similar to those obtained from the comparable inorganic buffers. 6. The effects of varying concentrations of potassium and sodium on the net absorption rate of Mg (as 24Mg) and on the one-way efflux of Mg (as 28Mg) from supernatant fractions or rumen fluid and inorganic buffers were investigated using the temporarily-isolated and washed rumen in three sheep. Although the net absorption rate of 24Mg from supernatant fractions or buffers containing similar K concentrations varied significantly between sheep, a similar percentage decrease in the absorption rates of both 24Mg and 28Mg was found for each sheep as the K concentration was increased. 7. One pair of sheep was fed on the frozen grass and the other pair was fed on the grass-maize pellets. Their daily intakes of K were then increased to 50 g/d for 14 d by intrarumen infusion of potassium chloride. In three of the four sheep the plasma Mg concentration fell within 12 h of the start of the KCl administration.(ABSTRACT TRUNCATED AT 400 WORDS)