Protection of leaf protein of lucerne (Medicago sativa L.) against degradation in the rumen by treatment with formaldehyde and glutaraldehyde

1980 ◽  
Vol 95 (3) ◽  
pp. 603-617 ◽  
Author(s):  
J. L. Mangan ◽  
D. J. Jordan ◽  
Janet West ◽  
P. J. Webb
Keyword(s):  
Medicago Sativa ◽  
Rumen Fluid ◽  
Major Effect ◽  
Toxic Effects ◽  
Leaf Protein ◽  
Tween 20 ◽  
Total N ◽  
Mean Values ◽  
Soluble Leaf Protein ◽  
Micro Organisms

SUMMARYAqueous glutaraldehyde, in the presence of wetting agents Tween-20 or Haemosol, reacted with fresh cut lucerne (Medicago sativa L.), complete reaction being obtained with about 7·2 g (72 mmol)/kg herbage, or 18g/100g crude protein. Reaction with 25% w/v aqueous glutaraldehyde sprayed on to fresh lucerne was rapid, and at the rate of 66 mmol/kg lucerne, all aldehyde had reacted in 3 h and about 60% of the soluble leaf protein became insoluble. Formaldehyde at twice the molar concentration of glutaraldehyde was absorbed rapidly, but a longer time, up to 24 h, was required for the protein to become insoluble. Treatments with 22, 44 and 66 mmol glutaraldehyde/kg lucerne, and 44, 88 and 132 mmol formaldehyde/kg showed that reaction with leaf protein was approximately proportional to the amount of aldehyde. A major effect on the leaf cells was the fixation of chloroplasts, and intact fixed chloroplasts were isolated from treated lucerne with high protein: chlorophyll ratios of 5·8:1 to 9·5:1.Two varieties of lucerne, Kabul and Europe, pot-grown in a controlled environment cabinet, reacted rapidly when sprayed with glutaraldehyde and in 3 h soluble leaf protein was reduced from 30 to 16–17% of the total N. The plants rapidly lost water and the dry matter of the leaves rose to 42% for Kabul and 45% for Europe in 24 h. Stems showed little effect. Field spraying of lucerne with glutaraldehyde similarly fixed soluble leaf protein and caused desiccation of the leaves, rising to 47–50% D. M. in 3 days. The stems were little affected and subsequent regrowth of the plants was not inhibited.Feeding glutaraldehyde- and formaldehyde-sprayed lucerne to rumen-fistulated cattle showed that release of soluble leaf protein into the rumen fluid was greatly reduced, mean values being 40 and 43% respectively of the values obtained when control lucerne was fed. Mean ammonia concentrations were similarly reduced to 49 and 33% of the control values. Formaldehyde-treated lucerne, even after reaction for several days, frequently showed toxic effects on rumen micro-organisms, particularly protozoa. Glutaraldehyde reacted more rapidly with herbage and no toxic effects were observed. Both glutaraldehyde- and formaldehyde-treated lucerne were highly palatable to cattle.

scholarly journals Characteristics of the rumen proteolysis of fraction I (18S) leaf protein from lucerne (Medicago sativa L)

1981 ◽  
Vol 46 (1) ◽  
pp. 39-58 ◽  
Author(s):  
J. H. A. Nugent ◽  
J. L. Mangan
Keyword(s):  
Medicago Sativa ◽  
Volatile Fatty Acids ◽  
Rumen Fluid ◽  
Maximum Velocity ◽  
Leaf Protein ◽  
Bacterial Cells ◽  
Fraction I Protein ◽  
Protein Nitrogen ◽  
Wide Range ◽  
Fraction I

1. The rate of proteolysis of fraction 1 (18S) leaf protein in the rumen of sheep and cattle was affected by diet and the rate on fresh lucerne (Medicago sativa L) was three to nine times the rate on a hay + concentrate diet.2. Simultaneous rumen fermentations in vivo and in an artificial rumen showed that the rates of proteolysis of fraction I in vitro was approximately 80% of the rates in sheep.3. Using 14C uniformly-labelled fraction I protein at low concentrations, proteolysis exhibited 1st-order kinetics. Over a wide range of protein concentrations the velocity v. substrate concentration curve showed Michaelis–Menten characteristics typical of an enzyme-catalysed reaction. With rumen fluid from a hay + concentrate-fed sheep the maximum velocity was 2.6 mg protein nitrogen/1 per min and the Michaelis constant was 75 mg nitrogen/1.4. Rapid adsorption of 14C-labelled fraction I protein onto bacterial cells preceded proteolysis.5. Sucrose-density-gradient analysis showed initial incorporation of 14C from protein into rumen bacteria followed by partial transfer to rumen protozoa.6. No peptides were detected during proteoiysis showing that the rate-limiting step occurred during the initial stages of proteolysis. Only small amounts of free amino acids were released except for leucine, isoleucine, valine and ornithine, which showed significantly increased levels.7. Volatile fatty acids were the main 14C-labelied end products and were rapidly produced in descending concentrations: acetate > propionate > 3-methyl + 2-methyl butyrate > butyrate > isobutyrate > valerate.

scholarly journals The effect of condensed tannins in Lotus pedunculatus on the solubilization and degradation of ribulose-1,5-bisphosphate carboxylase (EC 4.1.1.39; Rubisco) protein in the rumen and the sites of Rubisco digestion

1996 ◽  
Vol 76 (4) ◽  
pp. 535-549 ◽  
Author(s):  
W. C. Mcnabb ◽  
G. C. Waghorn ◽  
J. S. Peters ◽  
T. N. Barry
Keyword(s):  
Condensed Tannins ◽  
Rumen Fluid ◽  
Condensed Tannin ◽  
The Other ◽  
Leaf Protein ◽  
Bisphosphate Carboxylase ◽  
Freeze Dried ◽  
Lotus Pedunculatus ◽  
Micro Organisms

Three experiments were undertaken to determine the effect of condensed tannin (CT) in Lotuspedunculutus (45–55 g extractable CT/kg DM) on the digestion of the principal leaf protein, ribulose-1,5-bis phosphate carboxylase EC 4.1.1.39; Rubisco; fraction 1 leaf protein). In two of the experiments Lotus pedunculutus was fed to sheep, with one group receiving a continuous intraruminal infusion (per fistulum) of PEG (molecular weight 3500) to bind and inactivate the CT (PEG group). The other group, which did not receive PEG, was termed the control sheep (CT acting). Expt 3 involved in vitvo incubations of Lotus pedunculutus in buffered rumen fluid, with and without PEG added. In all experiments the results have been interpreted in terms of the effects of CT on Rubisco solubilization and degradation. Disappearance of N and Rubisco from Lotus pedunculutus suspended in polyester bags in the rumen was used as a measure of solubilization. Degradation was defined as the disappearance of Rubisco from in vitro incubations of Lotus pedunculatusinrumen fluid. In Expt 1, CT reduced the digestion of Rubisco in the rumen from 0.96 to 0.72 of intake (P < 0.01). Rubisco digestion in the small intestine was 0.27 of intake in control sheep and 0.04 of intake in PEG sheep. In Expt 2, PEG had no effect on the loss of Rubisco from Lotus pehneulatus contained in polyester bags which were incubated in the rumen, hence CT did not affect the solubilization of Rubisco. Observations in Expt 1 were confirmed by in vitro incubations in Expt 3, where PEG addition substantially increased the rate of degradation of plant protein to NH2. Addition of PEG decreased the period of time taken to degrade 50% of the Rubisco from about 13.8 h to about 3.0 h. It was concluded that the action of CT reduced the digestion of Rubisco in the rumen of sheep fed on fresh Lotus pedunculutus, and that this was primarily due to the ability of CT to slow its degradation by rumen micro-organisms, without affecting its solubilization. Both fresh-minced, and freeze-dried and ground lotus were used for in saccoand in vitro incubations; however, fresh-minced lotus was more suitable for the evaluation of protein solubilization and degradation in fresh forages.

Alterations in Ruminal Fluid and Serum Biochemical Constituents in Goats Affected with Ruminal Acidosis

2018 ◽  
Vol 13 (03) ◽  
Author(s):  
P. R. Chavelikar ◽  
G. and Neha Rao C. Mandali ◽  
Neha Rao
Keyword(s):  
Rumen Fluid ◽  
Clinical Signs ◽  
Small Ruminants ◽  
Ruminal Fluid ◽  
Mean Values ◽  
Biochemical Constituents ◽  
Ruminal Acidosis ◽  
Methylene Blue Reduction ◽  
Serum Biochemical ◽  
The Mean

Ruminal acidosis is an important clinical emergency in small ruminants. In this study, eight healthy farm goats and 24 goats presented at TVCC of the college of Veterinary Sciences and A.H., Anand with clinical signs of ruminal acidosis and having rumen liquor pH below 6 were examined for alterations in the ruminal fluid and serum biochemical parameters. Among various rumen fluid parameters evaluated, the mean values of rumen fluid pH decreased significantly (4.71±0.11 vs. 6.90±0.10), while sediment activity time (46.67±1.20 vs. 24.50±0.78 min) and methylene blue reduction time (29.50±0.73 vs. 10.03±0.27 min) increased significantly in acidotic goats. The normal greenish, aromatic viscous color, odour and consistency of rumen fluid of healthy goats also changed to milky grey/creamy, sour/pungent watery in acidotic goats. The rumen protozoal activity decreased to nil in acidotic goats as compared to the healthy goats. Among various serum biochemical constituents, the mean values of glucose (92.43±1.37 vs. 74.13±1.83 mg/dl), BUN (26.49±0.47 vs. 22.63±1.19 mg/dl), serum creatinine (01.01±0.02 vs. 00.83±0.02 mg/dl) and albumin (03.22±0.03 vs. 03.05±0.05 g/dl), ALT (56.75±1.55 vs. 27.88±1.14 IU/L) and AST (93.25±1.82 vs. 54.00±1.75 IU/L), increased significantly, while there was significant decrease in serum calcium (09.09±0.14 vs. 10.29±0.08 mg/dl) in acidotic goats. The mean values of alkaline phosphatase (IU/L) in acidotic goats increased non-significantly from the base values of healthy goats.

The effect and site of action of potassium upon magnesium absorption in sheep

1976 ◽  
Vol 27 (6) ◽  
pp. 873 ◽  
Author(s):  
FM Tomas ◽  
BJ Potter
Keyword(s):  
Potassium Chloride ◽  
Rumen Fluid ◽  
Substantial Reduction ◽  
Urine Volume ◽  
Fluid Flows ◽  
Basal Diet ◽  
Major Effect ◽  
Urinary Potassium ◽  
Bioelectrical Potential ◽  
Plasma Magnesium

The effect of potassium chloride infusion to the rumen or duodenum of sheep upon the absorption of magnesium from the stomach and intestinal regions has been examined. Three Merino ewes, each prepared with a cannula into the rumen and a re-entrant cannula into the duodenum, were offered a basal diet (control) which provided 46.3–51.1 mmoles magnesium and 299–320 mmoles potassium per day. Potassium chloride (500–800 mmoles/day) was infused continuously to either the rumen or duodenum. Digesta fluid flows were estimated from the dilution of a Cr-EDTA solution continuously infused to the rumen. Potassium infusion to either gastrointestinal site led to a comparable increase in the water intake, urine volume and levels of plasma and urinary potassium. Infusion to the rumen caused a marked increase in the potassium levels and a decrease in sodium levels in rumen fluid, as well as an increase in the rumen fluid to blood bioelectrical potential. No effect of treatment on digesta fluid flows was observed. Net magnesium absorption was lowered only when potassium was infused to the rumen, and the reduction was almost entirely due to reduced absorption of magnesium from the stomach. There was no consistent effect on absorption of magnesium from the intestines. Plasma magnesium levels were lowered by both the intraruminal infusion and, to a lesser extent, the duodenal infusion of potassium. The results indicate that although one consequence of potassium ingestion by sheep may be an enhancement of the urinary excretion of magnesium, the major effect on magnesium metabolism is a substantial reduction of absorption of magnesium from the reticulorumen.

scholarly journals Hydrolysis of14C-labelled proteins by rumen micro-organisms and by proteolytic enzymes prepared from rumen bacteria

1983 ◽  
Vol 50 (2) ◽  
pp. 345-355 ◽  
Author(s):  
R. J. Wallace
Keyword(s):  
Proteolytic Enzymes ◽  
Rumen Fluid ◽  
Performic Acid ◽  
Cross Linking ◽  
Acid Oxidation ◽  
Rumen Bacteria ◽  
Rate Of Hydrolysis ◽  
Micro Organisms ◽  
Hydrolysis Of

1. Proteins were labelled with14C in a limited reductive methylation using [14C]formaldehyde and sodium borohydride.2. The rate of hydrolysis of purified proteins was little (< 10%) affected by methylation and the14C-labelled digestion products were not incorporated into microbial protein during a 5 h incubation with rumen fluid in vitro. It was therefore concluded that proteins labelled with14C in this way are valid substrates for study with rumen micro-organisms.3. The patterns of digestion of14C-labelled fish meal, linseed meal and groundnut-protein meal by rumen micro-organisms in vitro were similar to those found in vivo.4. The rates of hydrolysis of a number of14C-labelled proteins, including glycoprotein II and lectin from kidney beans (Phaseolus vulgaris), were determined with mixed rumen micro-organisms and with proteases extracted from rumen bacteria. Different soluble proteins were digested at quite different rates, with casein being most readily hydrolysed.5. Proteins modified by performic acid oxidation, by cross-linking using 1,6-di-iso-cyanatohexane or by diazotization were labelled with14C. Performic acid treatment generally increased the susceptibility of proteins to digestion, so that the rates of hydrolysis of performic acid-treated proteins were more comparable than those of the unmodified proteins. Cross-linking resulted in a decreased rate of hydrolysis except with the insoluble proteins, hide powder azure and elastin congo red. Diazotization had little effect on the rate of hydrolysis of lactoglobulin and albumin, but inhibited casein hydrolysis and stimulated the breakdown of γ-globulin.

Nitrogen excretion in germ-free and conventional chickens: effects of an alkali load

1978 ◽  
Vol 39 (1) ◽  
pp. 99-104 ◽  
Author(s):  
J. Okumura ◽  
D. Hewitt ◽  
Marie E. Coates
Keyword(s):  
Amino Acids ◽  
Uric Acid ◽  
Ammonia Excretion ◽  
Nitrogen Excretion ◽  
Acid Base Balance ◽  
Acid Base ◽  
Total N ◽  
Germ Free ◽  
Base Balance ◽  
Micro Organisms

1. Groups of three colostomized germ-free (GF) and conventional (CV) chickens aged 4 months were maintained for successive periods of 8 d on a diet containing 200 g casein/kg without and with sodium bicarbonate at the rate of 20 mmol/d and a nitrogen-free diet without and with NaHCO3at 9 mmol/d. Urine and faeces were collected during the last 3 d of each period.2. Total N, uric acid- and ammonia-N were determined in urine and total N in faeces. Amino acids were measured in hydrolysates of faeces collected during the periods when no NaHCO3was included in the diets.3. The CV birds excreted more N on the casein diets but less on the N-free diets than did their GF counterparts, the differences being mainly shown in the urine.4. On both diets hydrolysates of the faeces of CV birds contained smaller amounts of amino acids. On the N-free diet the proportions (g/160 g N) of serine, proline and threonine were reduced, suggesting some conservation of endogenous N by micro-organisms, and the proportions of histidine, alanine, lysine and methionine increased, possibly through microbial synthesis; on the casein diet, proportions of most amino acids were less, probably because bacterial deamination had occurred.5. Urinary excretion of total N, uric acid and ammonia was much greater on the casein than on the N-free diets. Inclusion of NaHCO3caused a sharp fall in urinary ammonia on both diets and in both environments.6. It was concluded that the level of dietary protein and the regulation of acid-base balance have more effect than microbial activity on the urinary ammonia excretion.

scholarly journals N-6 and n-3 fatty acids in different beef adipose tissues depending on the presence or absence of the gene responsible for double-muscling

2008 ◽  
Vol 53 (No. 12) ◽  
pp. 515-522 ◽  
Author(s):  
N. Aldai ◽  
M.E.R. Dugan ◽  
A.I. Nájera ◽  
K. Osoro
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Intramuscular Fat ◽  
Mean Value ◽  
Adipose Tissues ◽  
Total N ◽  
Mean Values ◽  
Carbon Chains ◽  
Subcutaneous Adipose ◽  
Fatty Acid Compositions

Levels of n-6 and n-3 PUFAs, including those with 20 and 22 carbon-chains, in concentrate-fed Asturiana de los Valles (AV) yearling bulls with and without the double muscling gene <I>(mh/mh</I> = 24, <I>mh</I>/+ = 26, +/+ = 25) were measured to examine if this gene influences the pattern of PUFA deposition in different adipose tissues. Fatty acid compositions of muscle tissue (<I>longissimus thoracis</I>) and intermuscular and subcutaneous adipose tissues were determined by gas chromatography. The composition of intramuscular fat was unique compared to the other two adipose tissues which were similar in composition. In general, n-6 and n-3 fatty acid elongation and desaturation products were affected by AV genotype and this effect was most evident in n-3 PUFAs of the intramuscular fat of <I>mh/mh</I> (n-6/n-3 = 11.8 and 18:2n-6/18:3n-3 = 25.3) compared to <I>mh</I>/+ and +/+ animals (mean values of n-6/n-3 = 9.86 and 18:2n-6/18:3n-3 = 15.5). PUFA elongation and desaturation end products did not accumulate to any great extent in intermuscular and subcutaneous adipose tissues. Beef from <I>mh/mh</I> cattle showed greater deposition rates of n-3 elongation and desaturation products but their absolute content of total n-3 fatty acids was lower (21 mg/100 g meat) in comparison with <I>mh</I>/+ and +/+ cattle (mean value of 25 mg/100 g meat).

Repeatability of in vitro digestibility assays of forages when using fresh, frozen or freeze-dried cow faeces instead of sheep rumen liquor as sources of micro-organisms

1995 ◽  
Vol 1995 ◽  
pp. 110-110 ◽  
Author(s):  
S Akhter ◽  
E Owen ◽  
M K Theodorou ◽  
S L Tembo ◽  
E R Deaville
Keyword(s):  
Freeze Drying ◽  
Rumen Fluid ◽  
In Vitro Digestibility ◽  
Two Stage ◽  
Freeze Dried ◽  
Fresh Frozen ◽  
Sheep Rumen ◽  
Micro Organisms

Previous studies (El Shaer, Omed and Axford, 1987; Akhter, Owen, Fall, O'Donovan and Theodorou, 1994) with the two-stage in vitro procedure of Tilley and Terry (1963) have shown a high correlation between digestibilities of forages as determined using either sheep rumen liquor, sheep faeces or cow faeces as the microbial inoculum. In the first study of the of the present investigation one objective was to examine the repeatability of these digestibility measurements when made on different occasions. A second objective was to assess whether the correlations between faecal and rumen fluid based inocula could be improved if microorganisms were obtained from pairs rather than individual animals. The objective in the second study using forages of known in vivo digestibility, was to investigate the effect of freezing or freeze-drying of faeces on the repeatability of digestibilities of forages determined in vitro using micro-organisms from cow faeces.

On the role of higher plant and microbial lipases in the ruminal hydrolysis of grass lipids

1977 ◽  
Vol 38 (2) ◽  
pp. 225-232 ◽  
Author(s):  
R. M. C. Dawson ◽  
N. Hemington ◽  
G. P. Hazlewood
Keyword(s):  
Rumen Fluid ◽  
Higher Plant ◽  
Microbial Enzymes ◽  
Heat Treated ◽  
Ruminal Degradation ◽  
Complex Lipids ◽  
Microbial Lipases ◽  
Micro Organisms ◽  
Hydrolysis Of

1. The galactolipids of heat-treated, 14C-labelled rye grass S24 administered intraruminally to a sheep fed on an autoclaved diet were rapidly catabolized.2. When grass was homogenized with rumen contents devoid of higher plant lipases the grass galactolipids were rapidly metabolized, but were not metabolized when the rumen contents were boiled to destroy microbial galactolipases.3. 14C-labelled monogalactosyldiglyceride, digalactosyldiglyceride and triolein were metabolized, with the release of 14C-labelled fatty acids when incubated with a homogenate (100 g/l) of grass or clover in rumen fluid from a starved sheep, but not when the rumen fluid was heat-treated to destroy microbial enzymes.4. It is concluded that in the sheep the lipases of rumen micro-organisms play a major part in the ruminal degradation of ingested complex lipids of pasture.

In vitromicrobial growth and rumen fermentation of different substrates as affected by the addition of disodium malate

2005 ◽  
Vol 81 (1) ◽  
pp. 31-38 ◽  
Author(s):  
M. L. Tejido ◽  
M. J. Ranilla ◽  
R. García-Martínez ◽  
M. D. Carro
Keyword(s):  
Dry Matter ◽  
Microbial Growth ◽  
Rumen Fermentation ◽  
Gas Production ◽  
Mean Values ◽  
Batch Cultures ◽  
Production Values ◽  
Micro Organisms ◽  
Microbial N

AbstractThe effects of two concentrations of disodium malate on thein vitrofermentation of three substrates differing in their forage: concentrate ratio (0·8: 0·2, 0·5: 0·5 and 0·2: 0·8; g/g dry matter; low-, medium- and high-concentrate substrates, respectively) by rumen micro-organisms were studied using batch cultures. Rumen contents were collected from four Merino sheep offered lucerne hay ad libitum and supplemented daily with 400 g concentrate. Disodium malate was added to the incubation bottles to achieve final concentrations of 0, 4 and 8 mmol/l malate and15N was used as a microbial marker. Gas production was measured at regular intervals from 0 to 120 h of incubation to study fermentation kinetics. When gas production values were corrected for gas released from added malate, no effects (P> 0·05) of malate were detected for any of the estimated gas production parameters. In 17-h incubations, the final pH and total volatile fatty acid (VFA) production were increased (P< 0·001) by the addition of malate, but no changes (P> 0·05) were detected in the final amounts of ammonia-N and lactate. When net VFA productions were corrected for the amount of VFA produced from malate fermentation itself, adding malate did not affect (P> 0·05) the production of acetate, propionate and total VFA. Malate reduced methane (CH4) production by proportionately 0·058, 0·013 and 0·054 for the low-, medium- and high-concentrate substrates, respectively. Adding malate to batch cultures increased (P< 0·01) rumen microbial growth (mean values of 16·6, 18·3 and 18·4 mg of microbial N for malate at 0, 4 and 8 mmol/l, respectively), but did not affect (P> 0·05) its efficiency of growth (55·5, 56·7 and 54·3 mg of microbial N per g of organic matter apparently fermented for malate at 0, 4 and 8 mmol/l, respectively). There were no interactions (P> 0·05) malate × substrate for any of the measured variables, and no differences (P> 0·05) in pH, CH4production and microbial growth were found between malate at 4 and 8 mmol/l. The results indicate that malate had a beneficial effect on in vitro rumen fermentation of substrates by increasing VFA production and microbial growth, and that only subtle differences in the effects of malate were observed between substrates. Most of the observed effects, however, seem to be due to fermentation of malate itself.

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