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PubMed1972

Ruminal bacterial degradation of benzo(b)-thien-4-yl methylcarbamate (Mobam) and effect of Mobam on ruminal bacteria.

Mixtures of ruminal bacteria degraded benzo(b)thien-4-yl methylcarbamate (Mobam) to 4-hydroxybenzothiophene, CO(2), and polar product(s). The metabolite, 4-hydroxybenzothiophene, was identified (after acetylation) by comparative infrared and mass spectrometry with an authentic sample. Carbon dioxide and polar product(s) were produced by degradation of the methylcarbamate moiety. Ten previously characterized strains of ruminal bacteria with diverse physiological capabilities did not degrade Mobam. However, three tributyrin-hydrolyzing strains were isolated that did degrade Mobam. Mobam inhibited growth of two of ten strains isolated on Mobam-free glycerol-tributyrin enrichment medium. One of these strains was also sensitive to 2-carbomethoxy-propene-2yl dimethyl phosphate (Phosdrin). Mobam prevented some ruminal bacteria from producing zones of hydrolysis in tributyrin emulsion media and inhibited some ruminal bacteria from degrading 1-naphthyl acetate and fluorescein-3',6'-diacetate.

Anaerobiosis↗
PubMed1991

Influence of ruminal or duodenal soybean oil infusion on intake, ruminal fermentation, site and extent of digestion, and microbial protein synthesis in beef heifers consuming grass hay.

Six heifers (two Hereford X Jersey, four Hereford X Longhorn; average BW 278 kg) cannulated at the rumen and duodenum and fed a grass hay (fescue/orchardgrass) diet were used in a replicated 3 X 3 Latin square. Treatments were either no infusion (C), 150 ml of duodenally infused soybean oil (DI), or 150 ml of ruminally infused soybean oil (RI)/heifer twice daily for a total daily infusion of 300 ml of soybean oil. Periods of the Latin square included 18 d for adaptation and 5 d for collection. Forage OM, ADF, NDF, and N intakes were not affected (P greater than .10) by soybean oil infusion. Ruminal (P = .11) and total tract (P less than .10) OM digestibilities were decreased by RI compared with C or DI, but ADF and NDF digestibilities were not affected by treatment. Duodenal N (P less than .05) and microbial N flows were increased (P less than .10) for C and RI compared with DI. Microbial efficiency (g of N/kg of OM truly fermented) was improved (P less than .10) by RI compared with DI but did not differ (P greater than .10) from C. Ruminal pH was lower (P less than .05) with RI than with either C or DI. Ruminal NH3 N, total VFA, and acetate were not affected (P greater than .10) by treatment. Propionate (mol/100 mol) was greater (P less than .05) with RI than with DI and C, but the proportion of butyrate did not differ among treatments. These data indicate minimal direct benefits for improving forage usage as a result of soybean oil infusion with a 100% grass diet; however, animals should realize benefits from additional dietary energy provided by infused lipid.

Animal Feed↗
PubMed1993

Vitamin E is not degraded by ruminal microorganisms: assessment with ruminal contents from a steer fed a high-concentrate diet.

Using an in vitro incubation system containing undiluted ruminal contents from a steer fed a high-concentrate, corn-based diet, we examined microbial degradation of DL-alpha-tocopherol acetate (TA). Gas production, pH, and fermentation acid profiles were done in an initial experiment to ensure conditions for reproducible, viable cultures over 24 h. The pH decreased from 5.7 to 4.9, gas production averaged 3.4 mL/mL of ruminal contents, and > 300 mM fermentation acids were produced. We then monitored the fate of TA added to bottles containing ruminal contents. Three methods of TA extraction were tried, of which two were used in experiments. The two methods used were 1) hot ethanol in a Soxhlet apparatus and 2) chloroform/methanol. Each of these was used to extract added TA from a set of three in vitro experiments. Concentrations of TA were determined at 0 h and after 4, 8, and 24 h at 39 degrees C. In the three hot ethanol extracted experiments, TA recoveries were 85% at 0 h. With time of incubation, TA levels either 1) remained constant, 2) decreased then returned to the initial value, or 3) decreased by approximately 50%. These inconsistent results indicated that this extraction method was unacceptable. In the latter three experiments we used a chloroform/methanol extraction method. Recoveries of added TA averaged 96% overall. Thus, the level of TA remained constant during the 24-h period, suggesting that microbial destruction of TA does not occur. Rather, the previously reported losses of vitamin E may be attributable to incomplete extraction of tocopherol from high-concentrate ruminal contents.

Analysis of Variance↗
PubMed1993

Technical note: ruminal vein catheterization and continuous blood flow measurement in ruminal arteries of sheep.

Eight wethers were used to test the technique. Silicone rubber catheters were introduced into both ruminal veins so that their tips lay a few centimeters from the splenic vein. Arterial blood flow to the rumen was measured by an ultrasonic transit-time flow meter with 3-mm probes implanted around the left and right ruminal arteries. No loss of patency of the venous catheters was observed before slaughter (2 to 6 mo after surgery). There was no evidence of extensive vascular trauma due to catheterization at postmortem examination. In vivo calibration of the flow probes showed that reliable measurements could be made until at least 6 mo after implantation. With an accurate method of blood flow measurement in ruminal arteries and guaranteed long-term catheter patency, it would be possible to make reliable estimates of nutrient uptake across the ruminal wall of sheep over an experimental period of several months.

Animals↗
PubMed1986

Ruminal evacuation's effect on microbial activity and ruminal function.

The influence of evacuating, mixing and returning ruminal contents on microbial populations, volatile fatty acid (VFA) concentrations and liquid flow rate was investigated with four ruminal-cannulated Hereford steers (247 kg avg wt). Ruminoreticular contents were sampled, then completely removed, mixed for 5 min and returned to the rumen. Subsequent samples were taken immediately, 1 h and 4 h later. Non-evacuated steers were sampled at identical time intervals either 1 d before or after evacuation. Averaged over time, there was no significant difference between evacuated and non-evacuated steers in total anaerobic, cellulolytic and facultative bacteria, protozoa, oxidation-reduction potential, VFA concentrations, and liquid flow rates. There were no treatment X time interactions and, except for holotrich protozoa and VFA, no differences from time of sampling. Ruminal evacuation does not appear disruptive to anaerobiosis or detrimental to ruminal microorganisms and digestive processes.

Animals↗
PubMed1998

[Systemic effects of ruminal acidosis following ruminal drinking in dairy calves. A retrospective analysis of 293 cases].

In a retrospective study the data of 293 young calves (age upon admission less than four weeks) with evidence of ruminal drinking (rumen pH < 6.0) were analyzed for possible associations between ruminal acidosis and various parameters of systemic acid-base metabolism (blood pH, blood levels of HCO3- and L-lactate, as well as anion gap). On the basis of the degree of ruminal acidosis (pH < 5.0/ > or = 5.0) and evidence of diarrhea, the animals were assigned to one of four groups. The results indicate that severe ruminal acidosis leads to disturbances of systemic acid-base metabolism in young calves, too. With additional diarrhea, those disturbances can be complex.

Acid-Base Equilibrium↗
PubMed2004

Prevalence and distribution of peste des petits ruminants virus infection in small ruminants in India.

Peste des petits ruminants (PPR) is an acute febrile viral disease of goats and sheep characterised by mucopurulent nasal and ocular discharges, necrotising and erosive stomatitis, enteritis and pneumonia. The disease is endemic in India and causes large economic losses each year due to the high rates of mortality and morbidity in infected sheep and goats. The present study reports observations from 58 laboratory confirmed outbreaks of PPR and provides details of the prevalence of antibodies to PPR virus (PPRV) in 4,407 serum samples of small ruminants. Most of the clinical specimens used for the study originated from the northern and central parts of India. Serum samples used for the detection of antibodies to PPRV were derived from a greater number of regions within the country, however, these samples may not be a true representation of the target population (unvaccinated sheep and goats over 3 months old). Indigenously developed monoclonal antibody-based diagnostic kits were used for the detection of PPRV antigen (sandwich enzyme-linked immunosorbent assay [ELISA]) and antibody (competitive ELISA). Findings suggested that the disease outbreaks were more severe in goats than sheep and that the frequency of disease outbreaks was greater between the months of March and June (51.7%) as compared to other periods of the year. Based on the screening of the 4,407 sera samples, the antibody prevalence of PPRV in small ruminants in India was 33% (95% confidence interval: 32.3% to 33.7%). The prevalence of antibodies to PPRV was noted to differ between species (i.e. sheep versus goats), age groups and geographical regions. A greater proportion of the sheep (36.3%) versus the goat (32.4%) population was infected with PPRV. The distribution and prevalence of antibodies to PPRV among various age groups of animals indicated that goats were exposed at an earlier age than the sheep, suggesting that goats may be more susceptible to infection with PPRV. A greater number of positive cases were observed in the southern and southwestern part of the country (30%-60%) as compared to northern India (10%-30%). These findings may be correlated with variations in the sheep and goat husbandry practices within different geographic regions, the topography of different states and the socio-economic status of individual Indian farmers.

Age Factors↗
PubMed1992

Effect of coculture of anaerobic fungi isolated from ruminants and non-ruminants with methanogenic bacteria on cellulolytic and xylanolytic enzyme activities.

Neocallimastix strain N1, an isolate from a ruminant (sheep), was cocultured with three Methanobacterium formicicum strains, Methanosarcina barkeri, and Methanobrevibacter smithii. The coculture with Methanobacterium formicicum strains resulted in the highest production of cellulolytic and xylanolytic enzymes. Subsequently four anaerobic fungi, two Neocallimastix strains (N1 and N2) from a ruminant and two Piromyces species from non-ruminants (E2 and R1), were grown in coculture with Methanobacterium formicicum DSM 3637 on filter paper cellulose and monitored over a 7-day period for substrate utilisation, fermentation products, and secretion of cellulolytic and xylanolytic enzymes. Methanogens caused a shift in fermentation products to more acetate and less ethanol, lactate and succinate. Furthermore the cellulose digestion rate increased by coculture. For cocultures of Neocallimastix strains with Methanobacterium formicicum strains the cellulolytic and xylanolytic enzyme production increased. Avicelase, CMCase and xylanase were almost completely secreted into the medium, while 40-60% of the beta-glucosidase was found to be cell bound. Coculture had no significant effect on the location of cellulolytic and xylanolytic enzymes.

Anaerobiosis↗
PubMed2006

Cloning and characterization of an alternative transcript of ovine glucose 6-phosphate dehydrogenase gene: comparative approach between ruminant and non-ruminant species.

Glucose 6-phosphate dehydrogenase (G6PD) plays an important role in ruminant's lipogenesis, as it provides necessary compounds of NADPH for the synthesis of fatty acids catalyzing the first committed reaction in the pentose phosphate pathway. In this work the full length ovine glucose 6-phosphate dehydrogenase cDNA was isolated using a polymerase chain reaction based strategy. Two isoforms (OG6PDA and OG6PDB) were detected encoding a protein of 515 and 524 amino acids, respectively. Both deduced amino acid sequences reveal a well conserved protein containing all the important residues for its catalytic role. The extra nine amino acids encoded by OG6PDB cause a frameshift in the polypeptide chain resulting in changes around the area of the potential substrate binding site. A three-dimensional model of ovine G6PD protein shows that this frameshift cause structural changes in the catalytic binding "pocket" of the molecule. Southern blot and RT analysis revealed that ovine G6PD appears as a single copy gene while it is expressed, with slight variability, in all tissues analyzed. Moreover, expression analysis of the ovine G6PD isoforms showed that OG6PDB is expressed only in tissues where lipogenesis is high in ruminants. Thus, we hypothesize that in ruminants G6PD may be regulated by the ratio of the two transcripts, according to the existence stimulus.

Alternative Splicing↗
PubMed2000

Economics of prophylaxis against peste des petits ruminants and gastrointestinal helminthosis in small ruminants in north Cameroon.

Data on reproduction and mortality were collected over one year from 5100 sheep and 13,300 goats in treated and control flocks. The treated animals received vaccination against peste des petits ruminants (PPR) and anthelmintics twice a year. Productivity parameters (fecundity and mortality rates) obtained with and without prophylaxis were fitted into a benefit-cost economic analysis model and run for project lifespans varying from one to five years. At a 7% discount rate, the overall benefits for a project lifespan of five years were estimated as over 15 million FCFA and 11 million FCFA for sheep and goats, respectively. The benefit-cost ratio ranged from 2.26 to 3.27 in goats and 3.01 to 4.23 in sheep, depending on the project lifespan. It was concluded that PPR and gastrointestinal helminthosis are important causes of economic losses in small ruminants in Cameroon. A national or even a regional vaccination campaign against PPR and strategic anthelmintic treatment of small ruminants are recommended.

Animals↗
PubMed1975

Relationship between ruminal ammonia and nonprotein nitrogen utilization by ruminants. II. Application of published evidence to the development of theoretical model for predicting nonprotein nitrogen utilization.

Results from published experiments dealing with several aspects of nitrogen utilization by ruminants were used to test the concept of zero utilization of nonprotein nitrogen under conditions where more ammonia (greater than 5 mg ammonia nitrogen/100 ml) is in the rumen than can be converted to microbial protein. Results from experiments where the flow of non-ammonia nitrogen to the abomasum of sheep was measured indicate that when urea was the source of supplemental nitrogen, a constant quantity of amino acids reached the abomasum for all rations ranging from 10 to 23% crude protein. From growth studies, addition of nonprotein nitrogen to low protein, high energy rations caused an improved rate of gain. Additions of nonprotein nitrogen to rations resulting in predicted ruminal ammonia concentrations greater than 5 mg ammonia nitrogen/100 ml rumen fluid were without benefit. From lactation studies, nonprotein nitrogen supplementation did not improve milk production if the ration contained more than 12.5% crude protein prior to supplementation or if the predicted ruminal ammonia concentration was greater than 4 mg ammonia nitrogen/100 ml rumen fluid. The importance of the amino acid requirement of the animal as well as the composition of the ration in designing and evaluating nitrogen supplementation studies is discussed.

Ammonia↗
PubMed1992

[The diagnosis of Listeria encephalitis in ruminants using cultural and immunohistological techniques. I. Comparison of different selective media and culture techniques for the detection of Listeria from ruminant brains].

The selective L-PALCAMY differential enrichment broth, the Listeria enrichment broth of the International Dairy Federation, Oxford Listeria selective agar, and PALCAM Listeria selective agar were comparatively examined in the cultural isolation of Listeria spp. from ten ruminant brains. The L-PALCAMY medium proved to be superior to the IDF broth in both selectivity and productivity for Listeria spp. in the brain samples, which were also contaminated with other bacteria. The Oxford and PALCAM agars corresponded in their productivity for Listeria spp. The latter, however, was more selective than the Oxford agar. Bacterial counts of up to 1.2 x 10(9) CFU/g of brain stem sample were made from Listeria monocytogenes (L.m.), and up to 6.2 x 10(4) CFU/g from Listeria innocua. A total of 164 brains from ruminants showing CNS disturbances and/or pathoanatomical CNS alterations were examined using L-PALCAMY medium, and Oxford and PALCAM agar. L.m. could be isolated from 29 of the brains, and Listeria innocua from five. Cultural isolation of both Listeria spp. occurred in one brain. Of 27 brains containing L.m., which were also examined using cold enrichment, L.m. was isolated in 59.3% of the cases with direct culture, in 81.5% of the cases using selective warm enrichment, and in 77.8% of the cases by means of selective cold enrichment. Five cases each were identified solely by cold or warm enrichment, respectively. In investigations of further 69 ruminant brains the number of brains shown to contain L.m. could be increased from seven to 13 by means of selective cold enrichment for three months.

Animals↗
PubMed2001

Ruminal methanogenesis as influenced by individual fatty acids supplemented to complete ruminant diets.

The objective of the present study was to investigate the effects of seven different pure fatty acids on rumen fermentation using the rumen simulation technique (RUSITEC). The fatty acids were supplied to a complete ruminant diet at a proportion of 50 g x kg(-1) dietary dry matter and compared with an unsupplemented control. Methane release and methanogenic counts were suppressed by the fatty acids C12 : 0, C14 : 0 and C18 : 2 whereas C8 : 0, C10 : 0, C16 : 0 and C18 : 0 showed no corresponding effects. Apart from C12 : 0 and C18 : 2, C8 : 0 and C10 : 0 also adversely affected ciliate protozoa suggesting independence from the methane-suppressing effect of medium-chain fatty acids (MCFA). Although MCFA but not C18 : 2 reduced ruminal fibre degradation, the influence on other fermentation traits remained low. In conclusion, the supply of certain fatty acids to ruminant diets seems to have the potential to reduce methane release.

Animal Feed↗
PubMed1993

[The effect of starch sources barley, corn and potatoes and their ration proportions on nutrient digestibility and energy utilization in ruminants. 2. Fractions of ruminally and postruminally digested nutrients in cattle].

The investigations of this paper are part of a complex research project to develop energetic feed evaluation within the Net Energy Fat System. The aim was to obtain new results to estimate the relation between place as well as kind of nutrient digestion and energetic utilization of rations in cattle. The ruminal nutrient digestibility was measured in adult oxen on feeding level 1.7 by means of duodenal reentrant cannula for 9 rations including the starch sources barley, maize and potatoes and with their energy parts of 50, 25 and 10%. The intake of starch ranged from 484 to 2573 g/animal.d and the amounts of ruminal and postruminal digested starch from 444 to 2336 as well as 10 to 284 g/animal.d. For the organic matter, starch, water soluble carbohydrates and N free residual substances high relative parts of ruminal digested from apparent digested nutrients were measured with values between 78 and 88, 83 and 98, 93 and 97 as well as 88 and 100% respectively.

Animal Feed↗
PubMed1980

The effects of diet on some hepatic enzyme activities in the pre-ruminant and ruminating calf.

Three mild-replacer diets containing different amounts of lipid with differing fatty acid compositions were fed to pre-ruminant calves. These diets did not affect the specific activities of the hepatic enzymes, involved in carbohydrate and lipid metabolism, which were studied. With the onset of rumination there was a general decrease in the activities of the glycolytic enzymes and glucose-6-phosphate dehydrogenase (EC 1.1.1.43) and an increase in the activity of acetyl CoA synthetase (EC 6.2.1.1). In the ruminating calf the specific activities of some of the enzymes of carbohydrate metabolism were substantially greater on the concentrate diet than on the pelleted dried grass. In the case of glucose-6-phosphate dehydrogenase, this difference attained statistical significance. This may be related to the amount of glucose absorbed as such from the small intestine in animals on the different diets.

Animal Feed↗
PubMed2000

Mathematical estimations of hyper-ammonia producing ruminal bacteria and evidence for bacterial antagonism that decreases ruminal ammonia production(1).

Mixed ruminal bacteria (MRB) from cattle fed hay produced ammonia from protein hydrolysate twice as fast as MRB from cattle fed mostly grain, and a mathematical model indicated that cattle fed hay had approximately four-fold more hyper ammonia-producing ruminal bacteria (HAB). HAB had a high maximum velocity of ammonia production (V(max)) and low substrate affinity (high K(m)), but simulations indicated that only large changes in V(max) or K(m) would cause a large deviation in HAB numbers. Some carbohydrate-fermenting ruminal bacteria produced ammonia at a slow rate (CB-LA), but many of the isolates had almost no activity (CB-NA). The model indicated that the ratio of CB-LA to CB-NA had little impact on HAB numbers. Validations based on predicted ratios of HAB, CB-LA and CB-NA over-predicted the specific activity of ammonia production by MRB, but co-culture incubations indicated that washed MRB from cattle fed grain could inhibit HAB. Because autoclaved MRB had virtually no effect on HAB and the incubations were always carried out at pH 7.0, the inhibition was not simply a chemical effect (e.g. low pH).

Journal Article↗
PubMed1991

In vivo metabolism of 2,2'-diaminopimelic acid from gram-positive and gram-negative bacterial cells by ruminal microorganisms and ruminants and its use as a marker of bacterial biomass.

Cells of Bacillus megaterium GW1 and Escherichia coli W7-M5 were specifically radiolabeled with 2,2'-diamino[G-3H]pimelic acid ([3H]DAP) as models of gram-positive and gram-negative bacteria, respectively. Two experiments were conducted to study the in vivo metabolism of 2,2'-diaminopimelic acid (DAP) in sheep. In experiment 1, cells of [3H]DAP-labeled B. megaterium GW1 were infused into the rumen of one sheep and the radiolabel was traced within microbial samples, digesta, and the whole animal. Bacterially bound [3H]DAP was extensively metabolized, primarily (up to 70% after 8 h) via decarboxylation to [3H]lysine by both ruminal protozoa and ruminal bacteria. Recovery of infused radiolabel in urine and feces was low (42% after 96 h) and perhaps indicative of further metabolism by the host animal. In experiment 2, [3H]DAP-labeled B. megaterium GW1 was infused into the rumens of three sheep and [3H]DAP-labeled E. coli W7-M5 was infused into the rumen of another sheep. The radioactivity contents of these mutant bacteria were insufficient to use as tracers, but the metabolism of DAP was monitored in the total, free, and peptidyl forms. Free DAP, as a proportion of total DAP in duodenal digesta, varied from 0 to 9.5%, whereas peptidyl DAP accounted for 8.3 to 99.2%. These data reflect the extensive metabolism of bacterially bound DAP within the gastrointestinal tracts of ruminant animals and serve as a serious caution to the uncritical use of DAP as a marker of bacterial biomass in the digesta of these animals.

Animals↗
PubMed1994

Influence of Yucca shidigera extract on ruminal ammonia concentrations and ruminal microorganisms.

An extract of the desert plant Yucca shidigera was assessed for its possible benefit in ruminal fermentation. The extract bound ammonia in aqueous solution when concentrations of ammonia were low (up to 0.4 mM) and when the extract was added at a high concentration to the sample (20%, vol/vol). The apparent ammonia-binding capability was retained after autoclaving and was decreased slightly following dialysis. Acid-precipitated extract was inactive. No evidence of substantial ammonia binding was found at higher ammonia concentrations (up to 30 mM). When Y. shidigera extract (1%, vol/vol) was added to strained rumen fluid in vitro, a small (6%) but significant (P < 0.05) decrease in ammonia concentration occurred, apparently because of decreased proteolysis. Inclusion of Y. shidigera extract (1%, vol/vol) in the growth medium of the rumen bacterium Streptococcus bovis ES1 extended its lag phase, while growth of Butyrivibrio fibrisolvens SH13 was abolished. The growth of Prevotella (Bacteroides) ruminicola B(1)4 was stimulated, and that of Selenomonas ruminantium Z108 was unaffected. Protozoal activity, as measured by the breakdown of 14C-leucine-labelled S. ruminantium in rumen fluid incubated in vitro, was abolished by the addition of 1% extract. The antimicrobial activities were unaffected by precipitating tannins with polyvinylpyrrolidone, but a butanol extract, containing the saponin fraction, retained its antibacterial and antiprotozoal effects. Saponins from other sources were less effective against protozoa than Y. shidigera saponins. Y. shidigera extract, therefore, appears unlikely to influence ammonia concentration in the rumen directly, but its saponins have antimicrobial properties, particularly in suppressing ciliate protozoa, which may prove beneficial to ruminal fermentation and may lead indirectly to lower ruminal ammonia concentrations.

Ammonia↗