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PubMed1992

Comparative growth rates of various rumen bacteria in clarified rumen fluid from cows and sheep fed different diets.

Pure cultures of strains of different species of rumen bacteria were grown in filter-sterilized rumen fluid supplemented with glucose, bicarbonate, and reducing agent (cysteine and sulfide). Growth rates were determined in a series of experiments. Strains of species most abundant in the rumen grew more rapidly than strains of less abundant bacteria. Ammonia, amino acids, and peptides increased growth rates to some extent, but the greatest stimulatory effect for less abundant bacteria was provided by other factors, present in yeast extract. Factors released from lysates of mixed rumen microbes stimulated growth, but their rate of release was slow. It was concluded that, besides energy and nitrogen sources, growth factors of an as-yet-undetermined nature probably play an important role in determining the predominance of different bacterial species in the rumen.

Animals↗

PubMed2003

Two-step freezing procedure for cryopreservation of rumen ciliates, an effective tool for creation of a frozen rumen protozoa bank.

The present study aimed at the long-term storage of rumen protozoa as living cells in liquid nitrogen. The two-step or interrupted slow freezing procedure was used to cryopreserve six of the dominant species of rumen ciliates isolated from monofaunated animals, Dasytricha ruminantium, Entodinium caudatum, Epidinium ecaudatum caudatum, Eudiplodinium maggii, Isotricha prostoma, and Polyplastron multivesiculatum. We optimized the first step in the interrupted slow freezing procedure, from the extracellular ice nucleation temperature to the holding temperature, and studied the effects of the cooling rates on survival. In addition to the nature of the cryoprotectant (dimethyl sulfoxide), the equilibration temperature and equilibration time (25 degrees C and 5 min, respectively), and the holding time at subzero temperature (45 min) recommended previously (S. Kisidayová, J. Microbiol. Methods 22:185-192, 1995), we found that a holding temperature of -30 degrees C, a cooling rate from extracellular ice nucleation temperature to holding temperature of between 1.2 degrees C/min and 2.5 degrees C/min, depending on the ciliate, and rumen juice as the freezing and thawing medium markedly improved the survival rate. Survival rates determined after 2 weeks in liquid nitrogen were 100% for Isotricha, 98% for Dasytricha, 85% for Epidinium, 79% for Polyplastron, 63% for Eudiplodinium, and 60% for Entodinium. They were not significantly modified after a period of 1 year in liquid nitrogen. Four of the five ciliate species cryopreserved for 8 months in liquid nitrogen successfully colonized the rumen when inoculated into defaunated animals. These results have made it possible to set up a bank of cryopreserved rumen protozoa.

Animals↗

PubMed1982

Effect of water temperature on rumen temperature, digestion and rumen fermentation in sheep.

Three studies were conducted to examine effect of water temperature on rumen temperature, digestion and fermentation in sheep. Four, 2-yr-old wethers were used in a Latin square design among four water temperature treatments: 0, 10, 20 and 30 C. Rumen temperature was depressed most by 0 C water followed by 10 C, 20 C and 30 C water. For 0, 10, 20 and 30 C water, respectively, 108, 96, 96 and 72 min were needed to reach initial rumen temperature. Water temperature had no significant effect on N balance or percentage dry matter digestibility, crude protein digestibility and crude fiber digestibility although lowest digestion coefficients were observed for the 0 C treatment. Numerical differences were observed in rumen pH or volatile fatty acid (VFA) and ammonia-N concentrations at the various water temperatures; however, the effects were not significant. Rumen pH was maximally depressed at 2 h postfeeding for 0 C water, 3 h for 10 C water, 4 h for 20 C water, and 3 and 4 h for 30 C water. By 4 h postfeeding, the 0 C water treatment produced the highest concentration of all VFA and ammonia-N; however, by 4 h not all of the VFA or ammonia-N concentrations had reached maxima for 10, 20 or 30 C water treatments. The 0 C water treatment produced the lowest concentrations of ammonia-N and VFA (except acetate) by 5 h postfeeding.

Animals↗

PubMed1994

Comparison of utilization of pectins from various sources by pure cultures of pectinolytic rumen bacteria and mixed cultures of rumen microorganisms.

Utilization of citrus, lucerne, apple and sugar beet pulp pectins by pure strains of rumen bacteria, Prevotella ruminicola, Lachnospira multiparus and Butyrivibrio fibrisolvens was compared. Additionally, the utilization of pectins by mixed rumen microorganisms was evaluated. The comparison was based on the depletion of galacturonic acid from medium, content of cellular protein in the cultures and the amount of end products of pectin fermentation in cell-free culture fluids. It was found that citrus pectin was utilized best; utilization of lucerne, apple and sugar beet pectins was dependent on the species of bacteria. P. ruminicola and B. fibrisolvens utilized polygalacturonic acid from sugar beet pectins better than that from apple or lucerne pectin, while L. multiparus was capable of significantly better utilization of lucerne pectin than pectin from sugar beet or apple. The source of pectin was less important for mixed cultures of rumen microorganisms than for pure cultures of rumen bacteria. The amount of fermentation products in the culture fluids supported the conclusion that citrus pectin was utilized better than others. Microbial protein content in the cultures was found to be a less sensitive indicator of pectin utilization than the remaining examined parameters. P. ruminicola strains and mixed cultures of rumen microorganisms were shown to have the highest ability to utilize pectins, L. multiparus-moderate, while the B. fibrisolvens strains utilized pectin the least.

Animals↗

PubMed1980

Kinetic parameters of lactate dehydrogenases of some rumen bacterial species, the anaerobic ciliate Isotricha prostoma and mixed rumen microorganisms.

A number of kinetic parameters of the lactate dehydrogenases of three rumen bacterial species (Peptostreptococcus productus, Propionibacterium acnes and Actinomyces viscosus), the rumen ciliate Isotricha prostoma and mixed rumen microorganisms (MRM) with respect to NADH, pyruvate, fructose-1,6-diphosphate (FDP) as well as the effects of several nucleotide phosphates were studied. Partially purified LDH of Peptostr. productus had the same kinetic parameters as in crude cell free extracts. Values for Km, determined by Michaelis-Menten kinetics with pyruvate as the substrate, were in the same range for all lactate dehydrogenases. After feeding a cow, changes in the apparent Km and Vmax values for NADH of the total LDH activity in MRM were followed. It is suggested that of the factors studied the ratio NADH/NAD(H) and ATP are the most important regulatory factors for the lactate dehydrogenases of mixed rumen microorganisms.

Actinomyces↗

PubMed2003

Possible quorum sensing in the rumen microbial community: detection of quorum-sensing signal molecules from rumen bacteria.

The bioluminescence assay using Vibrio harveyi BB170 was used to examine quorum-sensing autoinducer 2 (AI-2) activity from cell-free culture fluids of rumen bacteria. The assay showed that the culture fluids of four species of rumen bacteria, Butyrivibrio fibrisolvens, Eubacterium ruminantium, Ruminococcus flavefaciens, and Succinimonas amylolytica, contained AI-2-like molecules. Furthermore, homologues for luxS genes were detected in rumen fluids collected from three cows and in bacterial cells of P. ruminicola subsp. ruminicola and R. flavefaciens. These findings suggest that the quorum-sensing system mediated by AI-2 is present in the rumen.

Animals↗

PubMed1985

Utilization of low-quality roughage by Bos taurus and Bos indicus cattle. 2. The effect of rumen-degradable nitrogen and sulphur on voluntary food intake and rumen characteristics.

In a number of experiments voluntary food intake of three low-quality roughages, either alone or supplemented with rumen-degradable nitrogen and sulphur and minerals, was measured in Brahman (Bos indicus) and Hereford (Bos taurus) steers. The chaffed hays were Spear grass (Heteropogon contortus) (6.2 g N/kg organic matter (OM)), Pangola grass (Digitaria decumbens) (7.9 g N/kg OM), and Pangola grass (12.0 g N/kg OM). Rumen characteristics relating to rate of fluid outflow from the rumen were also determined. There was no significant difference between breeds in the dry-matter intakes of the unsupplemented diets which ranged from 11.3 to 17.8 g/kg body-weight (BW) by Herefords and from 11.8 to 16.1 g/kg BW by Brahmans. Supplementation of Spear grass with N and S significantly (P less than 0.05) increased intake by Herefords (24%) but not by Brahmans. When the lower-N Pangola grass was supplemented there was a significant increase in intake by both breeds with the magnitude of the response in Herefords (42%) (P less than 0.001) being greater than that in Brahmans (15%) (P less than 0.05). The intakes of both the supplemented Spear grass and the lower-N Pangola diets were significantly (P less than 0.05) greater by Herefords than Brahmans. There was no breed difference in intake when the higher-N Pangola grass was supplemented. Both breeds recorded an 8% intake response to supplementation, although the increase was only significant (P less than 0.05) in Herefords. The mean retention time of fluid in the rumen on the unsupplemented Pangola grass diet of lower N content was 12.7 h in Brahmans compared with 17.5 h in Herefords (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Ammonia↗

PubMed1978

[Behavior of the trace elements copper, zinc and manganese in bovine rumen. 1. Trace element content of different fractions of rumen fluid and the effect of copper sulfate administration].

Two adult, rumen-fistulated steers were fed, over four consecutive 3-week periods, Cu-deficient and normal hays with and without copper sulphate supplementation. Their rumen liquor levels of copper, zinc and manganese were determined correspondingly. The rumen liquor samples were split into 3 fractions by way of centrifugation, sodium dodecyl sulphate treatment of the sediment and filtration. The fractionation of the rumen liquor showed copper to be contained mainly in the water-soluble supernatant (68%), whilst zinc and manganese are more strongly linked to the microbial fraction (57 and 46%, resp.). The plant residue did not average but 9, 24 and 22% of copper, zinc and manganese, respectively. Copper sulphate supplementation to normal hay resulted in a significantly increased copper content in the supernatant and in the microbial fraction. At the same time, the zinc and manganese content values in the microbial fraction were found increased. However, the same copper sulphate supplementation to Cu-deficient hay was found to reduce highly significantly the copper level in the microbial fraction. It is supposed that the excessively high molybdenum content of this hay (2.5 ng/kg) has prevented any effect of the copper sulphate supplementation. The positive effects of copper sulphate supplementation on the zinc and manganese levels in the bacterial fraction were maintained even when feeding Cu-deficient hay rich in Mo.

Animal Feed↗

PubMed1978

[Behavior of trace elements copper, zinc and manganese in bovine rumen. 2. Passage rate and course of trace element concentration in rumen as influenced by copper sulfate administration].

In experiments on the distribution of copper, zinc and manganese in fractions of rumen liquor (Wetzel and Menke, 1977 (1)) changes in PEG and in trace element concentration during day-time (between 8 a.m. and 6 p.m.) were measured at 2-hour intervals. Rumen volume was 61 l and passage rate 108 and 128 l/day in the two animals used. Average daily passage was calculated to be 40 mg copper, 98 mg zinc and 371 mg manganese. These values were found to be 11% higher than copper intake and 44 and 68% lower than zinc and manganese intakes, respectively. The differences indicate a remarkable endogenous secretion of copper, and a high absorption of zinc and manganese within the rumen. Concentration (mcg/g dry matter) of copper, zinc and manganese in the bacterial fraction and in total rumen liquor decreased slowly in the first hours after feeding, reaching a minimum after 4 to 6 hours and showed a slight increase thereafter. After addition of copper sulphate the curves mostly became linear. The conclusion is drawn that 1.8 ppm copper in inorganic binding does not lead to changes in trace element concentration to an extent that might be harmful to bacterial fermentation.

Animals↗

PubMed1981

[Effects of animal age and feeding regime on the quantitative and qualitative characteristics of intensively fattened lambs. 2. Rumen volume and structure of rumen mucosa].

Lambs of the species Merino mutton sheep kept on a perforated floor received, from their 38th day of life, either fattening feed for lambs ad libitum or fattening feed for lambs ad libitum plus chopped dried green fodder ad libitum (quota 17% of the intake of dry matter). At intervals of three weeks 4 lambs of each group were slaughtered from their 40th to their 166th day of life. The determination of the physical rumen volume of one half of the animals did not show significant differences between the two feeding regimes. The rumen volume showed an approximately linear increase (B = 0,84) with the increasing age of the animals. Hyper- and parakeratotic changes of the epithelialis of the rumen were caused by both feeding regimes. They were distinctly stronger and earlier developed in animals which only received fattening feed for lambs. The measuring of the rumen villi showed significantly smaller villi in all age groups of those animals which had taken in dried lucerne green fodder as a supplement.

Age Factors↗

PubMed1985

[Studies on rumen fermentation with special reference to the volatile fatty acid production rate in ewes during reproduction. 2. Production rate of volatile fatty acids and rumen volume].

Methods and results of the measuring of the rumen volume and the passage rate as well as the dependence of the production of volatile fatty acids on the parameters mentioned are described. The correlative relations between rumen volume on the one hand and feed passage on the other hand to the production quotas are investigated. Passage rates and rumen volumes were determined with the help of the isotope dilution method. In the course of reproduction rumen volumes of 7.81, 6.41, 8.91, 7.11 in experiment 1 and 5.41, 5.81, 9.51 and 4.71 in experiment 2 were measured in the sequence early stage of gestation, late stage of gestation, lactation and the period between pregnancies. The average total production of volatile fatty acids within 12 hours amounted to 475 mmol, 265 mmol, 313 mmol and 351 mmol in experiment 1 and 155 mmol, 175 mmol, 271 mmol and 144 mmol in experiment 2 in the same sequence. The average passage duration of the feed through the digestive tract of the ewes was 32.4 h in experiment 1 and 33.6 h in experiment 2. The relatively high feed intake of the ewes resulted in a short passage duration and could be a cause of the low production quotas of the volatile fatty acids.

Animals↗

PubMed1993

[Comparative studies of the parameters of rumen fermentation and the digestibility of feed rations in cattle and sheep. 1. Parameters of rumen fermentation].

The results from 283 studies of rumen liquid, taken by means of oesophagus probe, from different species of cattle (ox, cow, male and female young cattle) and sheep (adult wether) are compared. In the rumen liquid the parts of volatile fatty acids (VFA), total VFA content, pH value and the content of NH3 and urea were analysed. In the cattle's rumen liquid the part of acetic acid is on average 2 Mol-% and the content of total VFA up to 28 mmol/l (middle of the cow--sheep comparisons) higher than in that of wether. The contents of NH3 and urea are lower in cattle than in wether. Relations between the amount of difference of characteristic rumen fluid numbers of cattle and sheep and the crude fibre content of the rations, the live weight of the young cattle and the nutrition level are considered.

Ammonia↗

PubMed1977

The rumen flagellate Piromonas communis: its life-history and invasion of plant material in the rumen.

The rumen flagellat Piromonas communis is the zoospore of a phycomycete fungus inhabiting the rumen. Zoosporogenesis was stimulated by a dietary component (the inducer), and inhibited by compounds affecting membrane structure and function, but not by inhibitors of protein synthesis. The zoospores showed taxis towards the tissues surrounding the inflorescence of Lolium perenne L. in the rumen, invading principally the stomata and damaged tissues. The zoospores germinated on this substratum and the rhizoids of the developing vegetative stage penetrated the tissue, taking up C14 from labelled plant material, which was incorporated into the fungal cells. The conditions for maximum flagellate production (39 degrees C, pH 6-0 to 7-0, high concentration of CO2, absence of O2) resembled those found in the rumen. The organism was cultured in an undefined medium in vitro in the absence of other flagellates.

Animals↗

PubMed2000

Impact of dietary rumen undegradable protein and rumen-protected choline on intake, peripartum liver triacylglyceride, plasma metabolites and milk production in transition dairy cows.

The objectives of the present study were to determine the effects of rumen undegradable protein (RUP) level of prepartum diets, the supplementation of a rumen-protected choline product, and their interactions on milk production, feed intake, body weight and condition, blood metabolites, and liver triacylglycerides in dairy cows. Rumen-protected choline (RPC) was fed with two levels of RUP to 48 multiparous Holstein cows in a 3 x 2 factorial arrangement of treatments. Beginning 28 d before expected calving, cows were fed 10% rumen degradable protein, either 0, 6, or 12 g/d of RPC as CapShure (Balchem Corp., Slate Hill, NY) and either 4.0 or 6.2% RUP. After calving and through 120 d of lactation, cows received a common diet and continued RPC as per their prepartum assignment. Prepartum dry matter intake (kg/d) was not affected by RPC or RUP. Postpartum intake decreased when 6.2% RUP was fed prepartum. Milk production to 56 d in milk was decreased when cows were fed 6.2% RUP prepartum. Milk protein (kg/d) decreased when additional RUP was fed prepartum. Cows fed RPC lost more weight during the study period and tended to lose more body condition. Plasma urea nitrogen levels in the prepartum period were reduced for cows fed 4.0% RUP prepartum. Mean liver triacylglyceride determined from samples obtained at -28, -14, +1, +28, and +56 d in milk was not affected by RPC, prepartum RUP, or their combinations. Feeding 12 g of RPC/d in conjunction with 4.0% RUP increased milk production, but feeding RPC with 6.2% RUP prepartum and through 56 d in milk decreased production. The data indicate that 6.2% RUP does not benefit close-up dry cows, and the response to RPC depends the RUP content of the prepartum diet.

Animal Nutritional Physiological Phenomena↗

PubMed1989

Effect of three concentrate feeding frequencies on rumen protozoa, rumen digesta kinetics, and milk yield in dairy cows.

Three rumen-cannulated Holstein cows in mid to late lactation were randomly assigned to a 3 x 3 Latin square with the following concentrate feeding frequencies: one, two, and four times daily. Cows were fed ad libitum a diet containing 55% corn silage and 45% concentrate (DM basis). The forage portion of the diet was fed twice daily to all cows separately from the concentrate. Each experimental period was 20 d in duration. After a 12-d adaptation, daily individual feed consumption and milk production were measured. Rumen digesta were collected prior to (0 h) and at 2, 4, and 8 h after the morning feeding to measure postprandial changes in protozoa numbers, pH, VFA, and NH3 N. Chromium-EDTA and ytterbium chloride were used as liquid and particulate markers to estimate liquid and solid digesta turnover rate in the rumen. Concentrate and forage DM intake, ruminal protozoa numbers, pH, VFA, and NH3 N concentrations, and digesta turnover rate were not affected by concentrate feeding frequency. There was a significant concentrate feeding frequency with sampling time interaction influence on NH3 N in which feeding concentrate four times daily minimized postprandial fluctuation of rumen NH3 N concentration. Feeding concentrate four times daily increased milk fat and protein production.

Animal Feed↗

PubMed2003

Population dynamics of rumen microbes using modern techniques in rumen enhanced solid incubation.

The microbial ecology of the rumen is very complex. Different species of bacteria, protozoa, and fungi are involved in digestion of plant material in ruminants. In spite of complicated interrelationships among the various groups of microorganisms in the rumen ecosystem, Bacteria and Archaea are believed to play a major role because of their numerical predominance and metabolic diversity. In this work we are presenting the results for microbial population dynamics of rumen microbes during two-stage anaerobic digestion of grass. The reactors were inoculated with fresh rumen content. Fluorescent in situ hybridization, confocal laser scanning microscopy and epifluorescence microscopy were employed for microbial investigation. It was observed that Bacteria dominated in the hydrolytic reactor (1st stage) whereas Archaea were predominant in the methanogenic reactor (2nd stage). The stability of the methanogenic reactor was result of the dominance of Methanosaeta species (mainly the filamentous type).

Animals↗

PubMed1983

[Effect of rumen ciliates on the digestive utilization of various carbohydrate-rich diets and on the end-products formed in the rumen. II. Utilization of inulin, saccharose and lactose].

Three diets rich in inulin, saccharose and lactose, respectively, were given to 10 rumen-fistulated sheep. Two animals were defaunated, two were inoculated with either Polyplastron multivesibulatum or Entodinium sp., and two others were inoculated with both. The latter two were bred in conventional conditions. All animals ingested the same amounts of carbohydrates in the three diets (21-22 g/kg P0.75/day). Dietary nitrogen content was similar (table 1). The ciliate population was improved with the inulin diet (fig. 1; table 2). With a mixed population, the Entodinium improved with the inulin diet (fig 1; table 2). With a mixed population, the Entodinium sp. genus was always predominant. Holotrich protozoa (mainly Isotricha) in the rumen of the conventional sheep represented 15 to 30% of the total ciliate biomass, indicating that they were able to metabolize these soluble sugars. We also observed that P. multivesiculatum can ferment cellulose and all the soluble carbohydrates proposed in these diets. However, Entodinium sp. development occurred mainly in the presence of the sugard produced during carbohydrate hydrolysis by other ciliates or bacteria. The highest organic matter digestibility, noted in faunated animals (table 3) was confirmed by the VFA concentration in the rumen (table 4). This could be explained either by an activation of bacterial metabolism due to predation or by the direct effect of ciliates on fermentations, or both. Modifications in the VFA composition varied with ciliate inoculation, showing that ciliate metabolism may vary with the nature of the energy in the diet or that the observed results depended on various opposite effects in which the intensity of each component was influenced by the diet. In general, the acetic acid molar proportion increased and propionic acid decreased when there was a considerable Entodinium sp. population. The effect on butyric acid was low with these diets. Higher ammonia and lactic acid concentrations were observed in the rumen of faunated than defaunated sheep, irrespective of the ciliate inoculum.

Animals↗

PubMed1994

[The relationship between the acidity of the rumen contents and the proliferation status of the rumen mucosa in cattle].

Two non pregnant, non lactating Jersey cows, fitted with rumen fistulas, were subjected to three feeding periods: high - low - high in energy. At the end of each period biopsies (approximately 1 cm2) were taken from the floor of the dorsal blind sack of the rumen for morphological examination. After in-vivo-isolation of the ventral sack of the rumen a mixture of volatile fatty acids (pH 5) has been infused and the time measured in which the pH value rose from 5 to 7. The ruminal mucosa reacted to the different energy levels of the ration, as expected, by proliferation or regression. With high mucosa the pH of the infused solution rose, at the maximum, 100% faster to pH 7 than with low mucosa. On that way the different absorption capacity of the ruminal mucosa as well as their importance for the regulation of the pH value in the rumen could be demonstrated.

Absorption↗