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Evaluation of a mathematical model of rumen digestion and an in vitro simulation of rumen proteolysis to estimate the rumen-undegraded nitrogen content of feedstuffs.

Twelve grain mixtures, one lucerne (Medicago sativa) hay and one maize silage which had been used in mixed diets for which dietary nitrogen undegraded in the rumen (UDN) had been estimated with duodenally-cannulated cows, were studied. Total N in the feeds was fractionated into pool A (N soluble in borate-phosphate buffer), pool B (total N--(pool A + pool C)) and pool C (acid-detergent-insoluble N or residual N after 24 h incubation in protease solution). N solubilization in protease solution containing 6.6 units/ml (substrate-saturating enzyme concentration) indicated the presence of subfractions in pool B, with different rates of solubilization. Such subfractions were not detectable from in situ, Dacron bag, estimates of N solubilization. UDN was estimated using a dynamic mathematical model and rate-constants obtained from N solubilization in protease solution or in situ. For three grain mixtures tested using the protease technique the model predicted UDN values of 7, 10 and 12% compared with values of 47, 66 and 59% estimated in vivo. The full range of experimental feeds was tested using the in situ technique and UDN values predicted by the model were used to derive UDN values for twelve mixed diets. The latter values were significantly but not closely correlated with those determined in vivo (r2 0.41, P less than 0.05). An attempt was made to simulate rumen proteolysis in vitro by choosing a protease enzyme concentration (0.066 units/ml) providing a proteolytic activity similar to that of whole rumen fluid. The experimental samples of feed were subjected to simulated rumen proteolysis for 18 or 48 h to resemble the mean retention times in the rumen for grain mixtures and roughages respectively. The residual N at the end of incubation was considered as an estimate of UDN. The UDN values estimated from simulated rumen proteolysis and those determined in vivo for twelve mixed diets were in close agreement (r2 0.61, P less than 0.01). Simulated rumen proteolysis can serve as a simple, rapid and sensitive method to estimate UDN in a variety of feedstuffs.

Animal Feed↗

Metabolism of aflatoxin, ochratoxin, zearalenone, and three trichothecenes by intact rumen fluid, rumen protozoa, and rumen bacteria.

The effect of rumen microbes on six mycotoxins (aflatoxin B1, ochratoxin A, zearalenone, T-2 toxin, diacetoxyscirpenol, and deoxynivalenol ) considered to be health risks for domestic animals was investigated. The mycotoxins were incubated with intact rumen fluid or fractions of rumen protozoa and bacteria from sheep and cattle in the presence or absence of milled feed. Rumen fluid had no effect on aflatoxin B1 and deoxynivalenol . The remaining four mycotoxins were all metabolized, and protozoa were more active than bacteria. Metabolism of ochratoxin A, zearalenone, and diacetoxyscirpenol was moderately or slightly inhibited by addition of milled feed in vitro. The capacity of rumen fluid to degrade ochratoxin A decreased after feeding, but this activity was gradually restored by the next feeding time. Ochratoxin A was cleaved to ochratoxin alpha and phenylalanine; zearalenone was reduced to alpha-zearalenol and to a lesser degree to beta-zearalenol; diacetoxyscirpenol and T-2 toxin were deacetylated to monoacetoxyscirpenol and HT-2 toxin, respectively. Feeding of 5 ppm (5 mg/kg) of ochratoxin A to sheep revealed 14 ppb (14 ng/ml) of ochratoxin A and ochratoxin alpha in rumen fluid after 1 h, but neither was detected in the blood. Whether such conversions in the rumen fluid may be considered as a first line of defense against toxic compounds present in the diet is briefly discussed.

Aflatoxin B1↗

Influence of intoxication with organophosphates on rumen bacteria and rumen protozoa and protective effect of clinoptilolite-rich zeolite on bacterial and protozoan concentration in rumen.

The effect of O-ethyl-S-(2-diisopropylaminoethyl) methylthiophosphonate on rumen bacteria and rumen protozoa was investigated in sheep (after premedication with clinoptilolite-rich zeolite and without that premedication). In control animals a decrease in the total concentration of rumen protozoa was observed 3-7 d after intoxication (particularly in small and large ones). In clinoptilolite-rich-zeolite-treated animals only a slight decrease in protozoan numbers occurred during the first hours after the intoxication. Similarly, in every category of rumen bacteria marked differences between the groups were recorded, particularly in concentration of lipolytic bacteria. The results suggest some protective effect of clinoptilolite-rich zeolite for rumen microbiota against the organophosphate poison.

Animals↗

Assessment of rumen processes by selected-ion-flow-tube mass spectrometric analysis of rumen gases.

This work investigated the potential to use measurement of the concentration of certain gases in the rumen headspace to gain information about rumen processes and as a potential diagnostic tool. We used new equipment (selected-ion-flow-tube mass spectrometer) that allows rapid and precise analysis of many of the gases present in a sample. Samples of rumen headspace gas and corresponding samples of rumen liquor were taken from three lactating cows, prepared with rumen fistulae, at intervals after receiving their morning feed allocation (grass silage and concentrates). Hydrogen sulfide, methyl sulfide, and dimethyl sulfide, were the predominant gases that were measured in the rumen headspace by this technique. The concentrations of these sulfur compounds declined over the interval after feeding, mirroring ammonia concentrations measured in rumen liquor, reflecting their common dependence on the fermentation of sulfur amino acids. Ammonia concentrations in rumen headspace gas varied in the opposite direction to the concentration of ammonia in rumen liquor and likely depend more on the pH of rumen liquor. Consideration of the pKa of ammonia suggests that ammonia concentrations in rumen gas will be very low below pH 6, representing a useful diagnostic for subacute ruminal acidosis. Low concentrations of volatile fatty acids were detected in rumen gas. The molar proportions of volatile fatty acids were similar in gas and liquor samples, with rumen gas containing slightly less acetic acid and disproportionately more valeric and caproic acids.

Acidosis↗

[Relationship between the volatile fatty acids (VFA) in the rumen and nitrogen secretion into isolated sheep's rumen].

Two sheep with a ruminal fistula and an isolated small rumen were studied for the secretion of ammonia nitrogen, urea nitrogen, and amino nitrogen into the isolated rumen at different levels of volatile fatty acids (VFA) (50, 133-97, and 97-66 M Mol 1(-1)) in the rumen. The VFA level in the rumen was found to exert a great influence on the quantitative secretion of endogenous nitrogen from the blood through the rumen wall into rumen content. When the VFA level in the rumen was increased by administration of a single dose of acetic, propionic, and butyric acid, the secretion of ammonia nitrogen and amino nitrogen abruptly dropped and the secretion of urea into the isolated rumen slightly increased. The over-all amount of nitrogen (NH3-N + urea-N + amino-N) that had passed into the isolated rumen in the course of an hour showed a highly significant correlation with the passage of nitrogen in the form of ammonia and amino nitrogen and was greatest before the application of VFA to the rumen, i.e. at the level of 50 m mol 1-1. Of the metabolites under study, which were passing to the isolated rumen, amino nitrogen shared the greatest proportion (45.38-46.54%). When the VFA level in the rumen was raised, the proportion of ammonia secreted to the isolated rumen decreased and the proportion of urea in the total amount of nitrogen increased.

Animals↗

Lysis of viable rumen bacteria in bovine rumen fluid.

Streptococcus bovis and Butyrivibrio sp. were labeled with thymidine-methyl-(3)H, washed, and resuspended in rumen fluid or rumen fluid fractions obtained from Holstein and Jersey cows fed alfalfa hay once daily. Factors affecting the lytic activity found in untreated rumen fluid were examined. Day to day variation and differences before and after feeding were observed for the same cow. There were also differences between cows on the same day. For a given rumen fluid, the rate of release of label was roughly proportional to the number of labeled cells present over a 100-fold range in concentration. Removal of protozoa largely abolished the lytic action of fresh rumen fluid for S. bovis, but some soluble lytic activity remained. Mixed rumen protozoa added to media containing labeled S. bovis caused label to appear in solution. In a sample of rumen fluid containing 4.3 x 10(4) protozoa/ml 5.2% of the S. bovis population were destroyed by protozoa per hr. The mean rate of destruction for 12 runs on whole rumen fluid was 8.7% per hr with a standard deviation of 6.05. Parallel experiments with Butyrivibrio indicated that soluble lytic factors were more important for this organism. They could be destroyed by autoclaving and were generated when viable rumen bacteria were resuspended in autoclaved rumen fluid. The lysis of S. bovis and Butyrivibrio, at equal cell densities, by mixed rumen protozoa was compared in 30% rumen fluid media, and Butyrivibrio appeared to be more readily lysed than S. bovis.

Animals↗

Effects of corn processing on growth characteristics, rumen development, and rumen parameters in neonatal dairy calves.

Neonatal Holstein calves were fed texturized calf starters containing 33% whole (WC), dry-rolled (DRC), roasted-rolled (RC), or steam-flaked (SFC) corn to investigate how corn processing method affects intake, growth, rumen and blood metabolites, and rumen development. In the first experiment, 92 Holstein calves (52 male and 40 female) were started at 2 +/- 1 d of age and studied for 42 d. Starter dry matter (DM) intake was measured and fecal scoring conducted daily. Growth and blood parameter measurements were conducted weekly. A subset of 12 male calves (3/treatment) was euthanized at 4 wk of age and rumen tissue sampled for rumen epithelial development measurements. Experiment 2 consisted of 12 male Holstein calves ruminally cannulated at 7 +/- 1 d of age. Rumen fluid and blood samples were collected during wk 2 to 6. In the first experiment, postweaning and overall starter and total DM intake were significantly higher in calves fed starter with DRC than RC or SFC. Postweaning and overall starter and total DM intake were significantly higher in calves fed starter with WC than SFC. Postweaning average daily gain was significantly greater in calves fed starter with DRC than SFC. Blood volatile fatty acid concentrations were significantly higher in calves fed starter with SFC than in calves fed all other treatments. Papillae length and rumen wall thickness at 4 wk were significantly greater in calves fed starter with SFC than DRC and WC, respectively. In experiment 2, calves fed starter with WC had higher rumen pH and lower rumen volatile fatty acid concentrations than calves fed all other starters. Rumen propionate production was increased in calves receiving starter with SFC; however, rumen butyrate production was higher in calves fed starter with RC. Results indicate that the type of processed corn incorporated into calf starter can influence intake, growth, and rumen parameters in neonatal calves. Calves consuming starter containing RC had similar body weight, feed efficiency, and rumen development but increased structural growth and ruminal butyrate production when compared with the other corn processing treatments.

3-Hydroxybutyric Acid↗

Rumen bacterial protein synthesis and the proportion of dietary protein escaping degradation in the rumen of sheep.

1. The effect of supplementing barley diets with urea (U), extracted, decorticated groundnut meal (GNM) or Peruvian fish meal (PFM) on rumen bacterial protein synthesis and the proportion of undegraded food protein passing to the duodenum of sheep has been examined. 2. Three wethers were given isonitrogenous, isoenergetic diets containing (g/kg dry matter (DM)): U 20, GNM 106 or PFM 78, the crude protein (nitrogen x 6.25) contents being 139, 145 and 148 respectively. The sheep were fed hourly, the mean daily intake of DM being 0.634 kg. 3. Rumen bacterial protein synthesis was determined using 35S and diaminopimelic acid (DAPA) as bacterial markers and polyethylene glycol (PEG) and chromic oxide as markers of digesta flow. Rumen volatile fatty acid (VFA) production rate was determined by a continuous infusion of [1-14C]acetate. 4. 35S and DAPA gave similar estimates of the proportion of bacterial N in the trichloroacetic acid-precipitable nitrogen of the rumen digesta, the mean value being 0.86. The VFA production rate did not vary significantly between diets, the mean being 5.8 mol/24 h. The flow of bacterial N from the rumen was calculated from the PEG and CR2O3 estimates of flow and the 35S and DAPA estimates of the proportion of bacterial N in the rumen. 35S and DAPA gave similar values (mean 12.5 g/24 h) and Cr2O3 gave a slightly lower value (11.5 g/24 h) than PEG (13.5 g/24 h). Dietary effects, averaged over the four methods, were not significant; the values were 13.0, 13.4 and 11.0 g/24 h for the U, GNM and PFM diets respectively. 5. Duodenal samples were taken from two 12 h continuous collections from re-entrant cannulas and the DM flow adjusted to total recovery of Cr2O3. The mean recovery Cr2O3 at the duodenum was 0.798. The rates of flow of DM were 0.296, 0.311 and 0.334 kg/24 h and of non-ammonia-N (NAN) 13.5, 15.2 and 15.4 g/24 h on the U, GNM and PFM diets respectively. 6. The concentrations of the essential amino acids in duodenal digesta were generally higher with the PFM diet than with either of the other two diets. The flow of most amino acids through the duodenum was generally higher on the PFM and GNM diets than on the U diet. 7. The energetic efficiency of bacterial protein synthesis was calculated to be 2.1 g bacterial N/mol VFA or 28 g bacterial N/kg organic matter fermented in the rumen. 8. From the estimates of bacterial N flow the rumen and NAN flow through the duodenum it was calculated that 0.22 and 0.69 of the supplemental N from GNM and PFM respectively passed through the rumen undegraded.

Amino Acids↗

Rumen motility in experimental acidosis of the rumen in sheep.

In rumen acidosis, induced by infusion of saccharose solution and solutions of different volatile fatty acids and lactic acid into the rumen, and during induced disturbances of acid-base equilibrium in the arterial blood the motility of the dorsal sac of the rumen, the pH of rumen content and the indices of acid-base equilibrium in the arterial blood were investigated. The pH and the indices were determined by the micromethod of Astrup with an Acid-Base-Cart ABC-1 unit. During saccharose-induced acidosis of the rumen the pH of its content was decreased and its motility was strongly inhibited. Acidification of rumen content corresponded to the dissociation constant of a given acid. The motility of the rumen was inhibited most strongly by butyric acid, followed with regard of this effect by acetic acid, propionic acid and lactic acid. It was found that hydrogen ions as well as anions and non-dissociated forms of acids produced in the rumen were responsible for inhibition of this motility and that the rise in the concentration of hydrogen ions in the blood had no inhibitory effect on the motility of the rumen.

Acetates↗

Effect of a high sulfur diet on rumen microbial activity and rumen thiamine status in sheep receiving a semi-synthetic, thiamine-free diet.

A semi-synthetic thiamine-free diet was used on weaned lambs to test the effect of a high sulfur level on the rumen, microbial activity and on the microbial production of thiamine. In vivo and in vitro kinetic studies, as well as the determination of the thiamine concentrations and thiaminase activity in the rumen, were performed during the 16 week experiment. A high sulfur level (0.6%) in the diet, in comparison with a normal sulfur level (0.2%), did not modify the microbial activity of the rumen with the exception of a slightly retarded decrease in the volatile fatty acid (VFA) rumen concentration. The rumen thiamine level and the thiaminase activity were not modified by the dietary sulfur level. In contrast, the rate of sulfate reduction into sulfide in the rumen increased progressively with the 0.6% sulfur diet. In conclusion, a high sulfur level (0.6%) in the diet of sheep did not modify the thiamine status of the rumen. It strongly increased the production of sulfides but an adaptation period of several weeks was required by the rumen microflora to reduce sulfate at a maximal rate.

Animals↗

A rumen linear programming model for evaluation of concepts of rumen microbial function.

A linear programming model provides for analysis of general input-output relationships in the rumen, for evaluation of competitive relationships among rumen microbes, and for computation of optimal relationships in the rumen. Eight rumen microbial groups defined on the bases of substrate specificity, nutrient requirements for growth, fermentation products, and relative metabolic activities comprise the central core of the model. Relative metabolic rates of microbial groups calculated from their cell sized were used as coefficients in the objective function. The model was used to evaluate effects of different amounts of protein from feed and various carbohydrates upon microbial population and fermentation patterns as accommodated by current concepts. During the several solutions of the model, considerable simplification of the rumen microflora occurred. This implies that current data and concepts, and the hypothesis regarding relative metabolic rate, as represented in the model, do not accommodate adequately competitions among the several rumen microbial species and, thus, that additional data and concepts regarding rumen microbial interactions are required. Also evaluated were effects of ingestion of bacteria by protozoa upon over-all rumen function, absolute microbial cell yields, cell yields per mole of adenosine triphosphate, and factors affecting these.

Adenosine Triphosphate↗

Intake limitations, feeding behavior, and rumen function of cows challenged with rumen fill from dietary fiber or inert bulk.

Twelve multiparous, rumen-cannulated cows (17 DIM) were fed 25 or 35% NDF diets without or with added rumen-inert bulk as water-filled plastic containers (500 ml each) within three 4 x 4 Latin squares (21-d periods). Added bulk equaled 25% of pretrial rumen volume for each cow. Objectives were to challenge cows with rumen fill in the form of dietary NDF and inert bulk to determine whether bulk affects intake similarly for diets of different NDF content or whether changes in behavior or rumen function allow cows to adapt to higher fill. Inert bulk had no effect on DMI for cows fed 25% NDF but decreased DMI for cows fed 35% NDF (18.7 vs. 16.6 kg/d). Volume of rumen digesta plus inert bulk, 102 L, was similar for 35% NDF treatments regardless of presence or absence inert bulk. Additional NDF or inert bulk increased the number of ruminating bouts per day, chewing time per unit of DMI or NDF intake, frequency of reticular contractions during rumination, and fractional passage rate of NDF from the rumen. Although these changes may help to alleviate fill, they were insufficient to maintain intake under conditions of high fill. Intake by cows receiving high fiber diets can be limited by reticulorumen capacity. Future study is required to determine the effect of rumen fiber digestion, fiber particle size, and animal characteristics on fill-limited intake.

Animal Feed↗

[The use of ribonucleic acids as markers for the measuring of microbial protein yield in the rumen. 2. The effect of sample treatment, the time of sampling and the composition of the ration on the RNA:N ratio in rumen microbes].

In two experiments the influence of the treatment of samples, the sampling time and the composition of the rations on the RNA: N ratio in the rumen microbes was checked. Experiment I proved that freezing (-21 degrees C), thawing and freeze-drying of isolated bacteria and protozoa from the rumen fluid and from the duodenal content did not result in a change of the RNA content and the RNA-N: total N relation. If, however, the rumen fluid is stored deep-frozen before the isolation of the bacteria the N content in the DM of the bacteria decreases by 17% and that of RNA by 30%. This results in a change of the RNA: N relation of 16%. In conclusion, the bacteria are to be isolated immediately after rumen fluid sampling. Isolated bacteria can be stored deep-frozen before RNA determination and then freeze-dried. Experiment II showed that the RNA content of the rumen protozoa varies according to the period after feeding. The RNA: N relation was 0.50, 0.92, 0.70 and 0.58 on average 0, 3, 6 and 8 h after feeding, in which the 3rd hour after feeding can obviously be considered the time of increased microbial activity. The conclusion from this variation is that more than one isolation of microbes must be carried out in the course of the day in order to achieve representative samples. These statements apply to easily and not easily fermentable protein as N source in the feed. It could also be proved that no essential variation is to be expected in the RNA: N relation in the microbes isolated from the rumen fluid in the range of 8-21% crude protein in the DM of the ration (roughage: concentrate = 55: 45). On average the rumen microbes contained 1.7 g RNA-N/16 g N, essential differences between bacteria and protozoa could not be ascertained. From the slight variation of the RNA-N: N relation in the isolated bacteria from various cows one can conclude that there is no need to isolate the microbes of each individual animal.

Animal Feed↗

Modeling of rumen water kinetics and effects of rumen pH changes.

Mechanistic elements for determining water kinetics and effects of pH on VFA production and cellulose hydrolysis were incorporated into a model of rumen digestion and metabolism. Elements necessary for water kinetics were estimates of salivary flow during eating, resting, and ruminating, time course and amount of drinking water, osmotic flux across the rumen wall based on changes in rumen fluid osmolality, and liquid turnover rate. Osmotic flux equations predicted a significant net absorption of water from rumens of dairy cattle. Water kinetic equations predicted experimental changes in rumen volume during twice daily feeding, and produced marker dilution data qualitatively resembling published data. Increased marker dilution during feeding was due to increased rumen volume. An empirical equation predicting rumen pH from VFA concentration was validated against literature data. Changes in molar percentages of acetate and propionate as pH declined below 6.2 were produced by decreasing cellulose hydrolysis and fermentation and by altering stoichiometric coefficients for fermentation of soluble sugars and starch. Insufficient data prevented an adequate challenge of equations predicting decreases in cellulose hydrolysis and changes in stoichiometric coefficients as pH decreased.

Animals↗