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PubMed1989

Inducible bacteriophages from ruminal bacteria.

The incidence of temperate bacteriophage in a wide range of ruminal bacteria was investigated by means of induction with mitomycin C. Supernatant liquid from treated cultures was examined for phagelike particles by using transmission electron microscopy. Of 38 ruminal bacteria studied, nine organisms (23.7%) representing five genera (Eubacteria, Bacteroides, Butyrivibrio, Ruminococcus, and Streptococcus) produced phagelike particles. Filamentous particles from Butyrivibrio fibrisolvens are the first of this morphological type reported from ruminal bacteria. All of the other particles obtained possessed polyhedral heads and long, noncontractile tails (group B-type phage). The limited range of morphological types produced by mitomycin C induction cannot yet account for the much wider range of types found in ruminal contents by direct examination. The presence of viral genetic material in a significant percentage of the bacteria tested, as well as in a range of different genera, indicates that viral genetic material may be a normal constituent of the genome of appreciable numbers of ruminal bacteria.

Animals↗
PubMed1990

Dynamics of ruminal ciliated protozoa in feedlot cattle.

Fluctuations in ciliated protozoan concentrations were monitored in 40 individually fed crossbred heifers that were stepped up to an 85% concentrate diet either slowly (12 days) or rapidly (3 days), with or without monensin (30 ppm). Ruminal fluid was withdrawn from all animals by stomach tube at the start of the study, after each group reached full feed, and at 14-day intervals thereafter throughout the finishing period until termination (day 119). Neither monensin nor speed of step-up affected (P greater than 0.10) total protozoan concentrations, ruminal pH, or lactic acid concentrations. Average protozoan concentrations peaked on day 5, progressively declined until day 56, and then increased (P less than 0.05), suggesting an adaptation to ruminal conditions. Concentrations of Isotricha spp. were higher (P less than 0.05) on the final two sampling dates than at any other time. After day 28, Entodinium, Isotricha, and Polyplastron were the only surviving genera. Protozoa were not detected in 11 heifers on day 42 and day 56, but only two animals were defaunated on day 119, indicating either exogenous or endogenous refaunation. Average protozoan concentrations were not different (P greater than 0.25) between ruminal samples collected by stomach tube the day before slaughter (2.8 x 10(5)/g) and digesta samples collected the next day (1.6 x 10(5)/g). In feedlot cattle, defaunation apparently is transitory and individual animals harbor a dynamic protozoan population that fluctuates in response to changing ruminal conditions.

Animal Feed↗
PubMed1992

Effects of antibacterial agents on in vitro ovine ruminal biotransformation of the hepatotoxic pyrrolizidine alkaloid jacobine.

Ingestion of pyrrolizidine alkaloids, naturally occurring plant toxins, causes illness and death in a number of animal species. Senecio jacobaea pyrrolizidine alkaloids cause significant economic losses due to livestock poisoning, particularly in the Pacific Northwest. Some sheep are resistant to pyrrolizidine alkaloid poisoning, because ovine ruminal biotransformation detoxifies free pyrrolizidine alkaloids in digesta. Antibacterial agents modify ruminal fermentation. Pretreatment with antibacterial agents may account for some animal variability in resistance to pyrrolizidine alkaloid toxicosis, and antibacterial agents can also be used for characterizing ruminal pyrrolizidine alkaloid-biotransforming microflora. The objective of this study was to evaluate the effects of antibacterial agents on biotransformation of a predominant S. jacobaea pyrrolizidine alkaloid, jacobine, in ovine ruminal contents. Ovine ruminal jacobine biotransformation was tested in vitro with 20 independent antibacterial agents. Low amounts of rifampin and erythromycin prevented jacobine biotransformation. Chlortetracycline, lasalocid, monensin, penicillin G, and tetracycline were slightly less effective at inhibiting jacobine biotransformation. Bacitracin, crystal violet, kanamycin, and neomycin were moderately inhibitory against jacobine biotransformation. Brilliant green, chloramphenicol, gramicidin, nalidixic acid, polymyxin B SO4, sodium azide, streptomycin, sulfisoxazole, and vancomycin had little to no effect on jacobine biotransformation. The antibiotics that were most effective at inhibiting biotransformation were those that are active against gram-positive bacteria. Therefore, gram-positive bacteria are most likely critical members of the jacobine-biotransforming consortia.

Animals↗
PubMed1992

Metabolism of toxic pyrrolizidine alkaloids from tansy ragwort (Senecio jacobaea) in ovine ruminal fluid under anaerobic conditions.

The ability of ovine ruminal fluid to metabolize pyrrolizidine alkaloid (PA) from Senecio jacobaea under anaerobic conditions was evaluated. Four fistulated sheep fed PA served as individual sources of ruminal fluid, which was incubated in a defined minimal salts medium under two different anaerobic conditions, denitrifying and methanogenic. Anaerobic cultures amended with ovine ruminal fluids (20%), PA (100 micrograms/ml), and a defined minimal salts medium were monitored for a period of several days. These cultures revealed that while PA was not depleted in sterile, autoclaved controls or under denitrifying conditions, it was metabolized during periods of active methanogenesis under methanogenic conditions. In addition, samples of ruminal fluid were separated by differential centrifugation under anaerobic conditions, and the resultant supernatants were tested for their ability to metabolize PA as compared with those of the respective uncentrifuged control fluids. Uncentrifuged controls exhibited a PA depletion rate of -4.04 +/- 0.17 micrograms of PA per ml per h. Supernatants 1 (centrifuged at 41 x g for 2 min), 2 (centrifuged at 166 x g for 5 min), and 3 (centrifuged at 1,500 x g for 10 min) exhibited significantly slower depletion rates, with slopes of data representing -1.64 +/- 0.16, -1.44 +/- 0.16, and -1.48 +/- 0.16 micrograms of PA metabolized per ml per h, respectively, demonstrating no statistically significant difference among the supernatant cultures. Microscopic evaluations revealed that protozoa were present in the control whole ruminal fluid and to a lesser extent in supernatant 1, while supernatants 2 and 3 contained only bacteria.(ABSTRACT TRUNCATED AT 250 WORDS)

Anaerobiosis↗
PubMed1993

Inhibition of ruminal cellulose fermentation by extracts of the perennial legume cicer milkvetch (Astragalus cicer).

Cicer milkvetch (Astragalus cicer L.) is a perennial legume used as a pasture or rangeland plant for ruminants. A study was undertaken to determine whether reported variations in its ruminal digestibility may be related to the presence of an antinutritive material. In vitro fermentation of neutral detergent fiber (NDF) of cicer milkvetch by mixed rumen microflora was poorer than was the fermentation of NDF in alfalfa (Medicago sativa L.). Fermentation of cicer milkvetch NDF was improved by preextraction of the ground herbage with water for 3 h at 39 degrees C. Such water extracts selectively inhibited in vitro fermentation of pure cellulose by mixed ruminal microflora and by pure cultures of the ruminal bacteria Ruminococcus flavefaciens FD-1 and Fibrobacter succinogenes S85. Inhibition of the cellulose fermentation by mixed ruminal microflora was dependent upon the concentration of cicer milkvetch extract and was overcome upon prolonged incubation. Pure cultures exposed to the extract did not recover from inhibition, even after long incubation times, unless the inhibitory agent was removed (viz., by dilution of inhibited cultures into fresh medium). The extract did not affect the fermentation of cellobiose by R. flavefaciens but did cause some inhibition of cellobiose fermentation by F. succinogenes. Moreover, the extracts did not inhibit hydrolysis of crystalline cellulose, carboxymethyl cellulose, or p-nitrophenylcellobioside by supernatants of these pure cultures of cellulolytic bacteria or by a commercial cellulase preparation from the fungus Trichoderma reesei. The agent caused cellulose-adherent cells to detach from cellulose fibers, suggesting that the agent may act, at least in part, by disrupting the glycocalyx necessary for adherence to, and rapid digestion of, cellulose.

Animals↗
PubMed1993

Estimation of ruminal bacteriophage numbers by pulsed-field gel electrophoresis and laser densitometry.

To investigate phage activity in the rumen, a method for quantifying phage has been developed. By differential centrifugation and ultrafiltration, phage particles were separated and concentrated from ruminal fluid. Linear double-stranded DNA from this fraction containing predominantly tailed phage was isolated and separated by size, using pulsed-field gel electrophoresis (PFGE). Laser densitometry of gel photographs allowed the numbers of phages with DNA in each size region to be calculated and, therefore, the total numbers per milliliter of ruminal fluid to be estimated. Phage numbers were estimated to be between 3 x 10(9) and 1.6 x 10(10) particles ml of ruminal fluid-1. The phage population, as gauged by the appearance of DNA on PFGE gels, had two major components. A broad region of DNA between 30 and 200 kb was always present on PFGE gels. It appears this region comprises DNA from a great many different phages and would include most of the temperate phages. In addition, discrete DNA bands ranging in size from 10 to 850 kb were frequently observed. DNA from one such band, of 12 kb in size, was shown to consist primarily of a single DNA type, suggesting that it originated from a specific phage. It is postulated that the discrete bands are due to epidemics or blooms of phage activity from specific, probably lytic, phages. The method that has been developed will greatly enhance future investigations into the interactions between the ruminal phage population, the ruminal bacterial population, and animal nutrition and growth.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1998

Assessment of reductive acetogenesis with indigenous ruminal bacterium populations and Acetitomaculum ruminis.

The objective of this study was to evaluate the role of reductive acetogenesis as an alternative H2 disposal mechanism in the rumen. H2/CO2-supported acetogenic ruminal bacteria were enumerated by using a selective inhibitor of methanogenesis, 2-bromoethanesulfonic acid (BES). Acetogenic bacteria ranged in density from 2.5 x 10(5) cells/ml in beef cows fed a high-forage diet to 75 cells/ml in finishing steers fed a high-grain diet. Negligible endogenous acetogenic activity was demonstrated in incubations containing ruminal contents, NaH13CO3, and 100% H2 gas phase since [U-13C]acetate, as measured by mass spectroscopy, did not accumulate. Enhancement of acetogenesis was observed in these incubations when methanogenesis was inhibited by BES and/or by the addition of an axenic culture of the rumen acetogen Acetitomaculum ruminis 190A4 (10(7) CFU/ml). To assess the relative importance of population density and/or H2 concentration for reductive acetogenesis in ruminal contents, incubations as described above were performed under a 100% N2 gas phase. Both selective inhibition of methanogenesis and A. ruminis 190A4 fortification (>10(5) CFU/ml) were necessary for the detection of reductive acetogenesis under H2-limiting conditions. Under these conditions, H2 accumulated to 4, 800 ppm. In contrast, H2 accumulated to 400 ppm in incubations with active methanogenesis (without BES). These H2 concentrations correlated well with the pure culture H2 threshold concentrations determined for A. ruminis 190A4 (3,830 ppm) and the ruminal methanogen 10-16B (126 ppm). The data demonstrate that ruminal methanogenic bacteria limited reductive acetogenesis by lowering the H2 partial pressure below the level necessary for H2 utilization by A. ruminis 190A4.

Acetic Acid↗
PubMed2002

Ammonia production by ruminal microorganisms and enumeration, isolation, and characterization of bacteria capable of growth on peptides and amino acids from the sheep rumen.

Excessive NH(3) production in the rumen is a major nutritional inefficiency in ruminant animals. Experiments were undertaken to compare the rates of NH(3) production from different substrates in ruminal fluid in vitro and to assess the role of asaccharolytic bacteria in NH(3) production. Ruminal fluid was taken from four rumen-fistulated sheep receiving a mixed hay-concentrate diet. The calculated rate of NH(3) production from Trypticase varied from 1.8 to 19.7 nmol mg of protein(-1) min(-1) depending on the substrate, its concentration, and the method used. Monensin (5 micro M) inhibited NH(3) production from proteins, peptides, and amino acids by an average of 28% with substrate at 2 mg/ml, compared to 48% with substrate at 20 mg/ml (P = 0.011). Of the total bacterial population, 1.4% grew on Trypticase alone, of which 93% was eliminated by 5 micro M monensin. Many fewer bacteria (0.002% of the total) grew on amino acids alone. Nineteen isolates capable of growth on Trypticase were obtained from four sheep. 16S ribosomal DNA and traditional identification methods indicated the bacteria fell into six groups. All were sensitive to monensin, and all except one group (group III, similar to Atopobium minutum), produced NH(3) at >250 nmol min(-1) mg of protein(-1), depending on the medium, as determined by a batch culture method. All isolates had exopeptidase activity, but only group III had an apparent dipeptidyl peptidase I activity. Groups I, II, and IV were most closely related to asaccharolytic ruminal and oral Clostridium and Eubacterium spp. Group V comprised one isolate, similar to Desulfomonas piger (formerly Desulfovibrio pigra). Group VI was 95% similar to Acidaminococcus fermentans. Growth of the Atopobium- and Desulfomonas-like isolates was enhanced by sugars, while growth of groups I, II, and V was significantly depressed by sugars. This study therefore demonstrates that different methodologies and different substrate concentrations provide an explanation for different apparent rates of ruminal NH(3) production reported in different studies and identifies a diverse range of hyper-ammonia-producing bacteria in the rumen of sheep.

Animals↗
PubMed2004

Effect of forage or grain diets with or without monensin on ruminal persistence and fecal Escherichia coli O157:H7 in cattle.

Twelve ruminally cannulated cattle, adapted to forage or grain diet with or without monensin, were used to investigate the effects of diet and monensin on concentration and duration of ruminal persistence and fecal shedding of E. coli O157:H7. Cattle were ruminally inoculated with a strain of E. coli O157:H7 (10(10) CFU/animal) made resistant to nalidixic acid (Nal(r)). Ruminal and fecal samples were collected for 11 weeks, and then cattle were euthanized and necropsied and digesta from different gut locations were collected. Samples were cultured for detection and enumeration of Nal(r) E. coli O157:H7. Cattle fed forage diets were culture positive for E. coli O157:H7 in the feces for longer duration (P < 0.05) than cattle fed a grain diet. In forage-fed cattle, the duration they remained culture positive for E. coli O157:H7 was shorter (P < 0.05) when the diet included monensin. Generally, ruminal persistence of Nal(r) E. coli O157:H7 was not affected by diet or monensin. At necropsy, E. coli O157:H7 was detected in cecal and colonic digesta but not from the rumen. Our study showed that cattle fed a forage diet were culture positive longer and with higher numbers than cattle on a grain diet. Monensin supplementation decreased the duration of shedding with forage diet, and the cecum and colon were culture positive for E. coli O157:H7 more often than the rumen of cattle.

Animal Feed↗
PubMed1962

Some nutritional characteristics of predominant culturable ruminal bacteria.

Bryant, M. P. (U.S. Department of Agriculture, Beltsville, Md.) and I. M. Robinson. Some nutritional characteristics of predominant culturable ruminal bacteria. J. Bacteriol. 84:605-614. 1962.-The effect of enzymatic hydrolysate of casein, NH(4) (+), a mixture of volatile fatty acids (acetic, n-valeric, isovaleric, 2-methylbutyric, and isobutyric), hemin, and ruminal fluid on growth of 89 freshly isolated strains of predominant culturable ruminal bacteria was studied, using basal media containing glucose, cellobiose, or maltose as energy source, minerals, cysteine, and S(=) as reducing agents, and H(2)CO(3)-HCO(3) (-) buffer. Of these strains, 13% (four morphological groups) grew poorly or not at all in defined medium plus casein hydrolysate; 6% (one morphological group) required casein hydrolysate; 56% (four morphological groups) grew with either NH(4) (+) or casein hydrolysate as the main source of nitrogen; and NH(4) (+), but not casein hydrolysate, was essential for 25% of the strains (five morphological groups). The volatile fatty acid mixture excluding acetate was essential for 19% of the strains (five morphological groups), and this mixture and acetate were necessary for good growth of 23% of the strains (one morphological group) when casein hydrolysate was excluded from the medium; 30% of the strains (one morphological group) required hemin. Similar studies are reported on 35 old laboratory strains of ruminal bacteria, most of which were previously identified. The results indicate that most strains of ruminal bacteria can be grown in defined media, and suggest the relative importance of NH(4) (+) and volatile fatty acids and the relative lack of importance of organic nitrogen compounds such as amino acids in the nutrition of these bacteria.

Amino Acids↗
PubMed2004

Anger response styles and blood pressure: at least don't ruminate about it!

BACKGROUND: Research on anger suggests a link with blood pressure (BP), but the findings are complex and highly variable; this is at least partly attributable to measurement issues. PURPOSE: In this study we used a new model of anger responding that comprises 6 independent anger response styles in 2 dimensions: Aggression, Assertion, Social Support Seeking, Diffusion, Avoidance, and Rumination. Linear and interactive relations between the anger response styles and resting and ambulatory BP were tested, controlling for traditional risk factors and level of hostility. METHODS: Data from 2 samples of different cardiovascular health status were examined. In Study 1, 109 healthy participants (45 men and 64 women) were recruited. Study 2 involved a sample of 159 hypertensive patients (90 men and 69 women). All participants provided demographic and health information; completed the Behavioral Anger Response Questionnaire, a hostility measure; and underwent resting BP measurement. Study 2 participants also provided 24-hr ambulatory BPs. RESULTS: Examination of linear effects revealed inconsistent associations between anger response styles and BP. The moderating effect of Rumination on the relationship between the other anger response styles and BP was examined next. Rumination had a deleterious influence on the relation between Avoidance and Assertion and resting and ambulatory BP levels. The moderating influence of Rumination on Social Support Seeking varied between the genders. CONCLUSIONS: Overall, the results suggest that rumination is a critical moderating variable in the relation of anger and BP.

Adaptation, Psychological↗
PubMed2000

On assessing individual differences in rumination on sadness.

The Rumination on Sadness Scale (RSS), an individual-difference measure of rumination on sadness, was developed as an alternative to the Ruminative Responses scale of the Response Styles Questionnaire (RRRSQ; Nolen-Hoeksema & Morrow, 1991). Research has shown the RRRSQ to consist of multiple, not highly intercorrelated factors; only I factor explicitly addresses rumination. In Study 1, a 1-factor solution to a principal components analysis was shown to hold for responses to the RSS. The RSS was also shown to be reliable. In Study 2, convergent and discriminant validity of the RSS were assessed. In Study 3, individuals with high RSS scores exhibited more distress regarding current concerns with the introduction of a delay period (to allow them to ruminate) after a sad mood induction.

Adolescent↗
PubMed1996

Postparturient change in endotoxin levels of ruminal fluid and serum in dairy cows.

The changes in the endotoxin levels of ruminal fluid and serum in postparturient cows were evaluated. Five cows were fed hay and concentrates on an individual basis (farm A) and the other 5 cows were given complete feed ad libitum (farm B) after parturition. Ruminal pH levels decreased in both groups after parturition. Subsequently, the ruminal endotoxin levels increased with the declining pH on both of the farms. The ruminal endotoxin levels were slightly higher in cows of farm B than those of farm A. Serum endotoxin levels also showed episodic fluctuations, however, there were no parallel changes between the endotoxin levels of the ruminal fluid and sera.

Animals↗
PubMed2006

Cognitive vulnerability to depression, rumination, hopelessness, and suicidal ideation: multiple pathways to self-injurious thinking.

In order to advance the detection and prevention of suicide, recent research has focused on predictors of suicidal ideation and behavior such as negative cognitive styles, dysfunctional attitudes, hopelessness, and rumination. In this study the relationships among these risk factors in the context of the Attention Mediated Hopelessness (AMH) theory of depression are examined. One hundred and twenty-seven undergraduates in the Cognitive Vulnerability to Depression (CVD) project were followed for 2.5 years. The CVD project followed initially nondepressed freshmen, at either high or low cognitive risk for depression, in order to predict onsets and recurrences of depressive disorders. The presence and duration of suicidal ideation were predicted prospectively by rumination and hopelessness, and hopelessness partially mediated the relationship between rumination and ideation and fully mediated the association between rumination and duration of suicidality. Further, rumination mediated the relationship between cognitive vulnerability and suicidal ideation.

Adult↗
PubMed1981

The relation of food quantity to rumination behavior.

Preliminary work suggested that the quantity of food ingested by retarded individuals who usually ruminated following meals was related to the frequency and duration of ruminating responses. This possible relation was experimentally examined by systematically varying food quantity from regular portions to satiation levels for three retarded individuals who exhibited high levels of ruminating. A clear functional relation of food quantity to ruminating emerged, with satiation procedures producing rapid and large decreases in the relatively high frequencies and durations of ruminating characteristic of baseline food quantity conditions.

Adult↗
PubMed1984

A parametric analysis of the relationship between food quantity and rumination.

Rumination is the chronic regurgitation, chewing, and reswallowing of previously ingested food. The study reported here, using a parametric design, examined the control of rumination by the quantity of food eaten at meals. The subjects were three profoundly retarded individuals who chronically emitted this behavior. The quantity of food by weight ingested daily was varied in 10-oz steps in both ascending and descending series (data were collected only after breakfasts and lunches). Ruminating decreased when food quantity increased and increased when food quantity decreased. In addition, there was a similar inverse interaction between breakfast food quantity and post-lunch ruminating. The data showed relatively rapid transitions in both frequency and duration at each meal size for all subjects. The data establish a clear functional relation between the quantity of food ingested and ruminating.

Journal Article↗
PubMed2006

Assessment of ruminal degradation, oral bioavailability, and toxic effects of anticoagulant rodenticides in sheep.

OBJECTIVE: To assess the rate and extent of ruminal degradation of warfarin, chlorophacinone, and bromadiolone in vitro and determine the oral availability and clinical and hemostatic effects of each anticoagulant rodenticide in adult sheep. ANIMALS: 3 Texel sheep. PROCEDURE: Samples of ruminal fluid were incubated with each of the anticoagulants to assess the kinetics of ruminal degradation over 24 hours. To determine the plasma kinetics of the anticoagulants, each sheep received each of the anticoagulants IV or via a rumenimplanted cannula at 2-month intervals (3 rodenticide exposures/sheep). At intervals during a 240- to 360- hour period after treatment, prothrombin time (PT) was measured, plasma anticoagulant concentration was assessed, and clinical signs of rodenticide poisoning were monitored. In plasma and rumen extracts, anticoagulant concentrations were determined via high-performance liquid chromatography. RESULTS: In the rumen extracts, anticoagulants were slightly degraded (< 15%) over 24 hours. In vivo, oral availability of warfarin, chlorophacinone, and bromadiolone was estimated at 79%, 92%, and 88%, respectively. Although maximum PT was 80 seconds after chlorophacinone and bromadiolone treatments, no clinical signs of toxicosis were detected; PT returned to baseline values within 2 weeks. CONCLUSIONS AND CLINICAL RELEVANCE: In sheep, warfarin, chlorophacinone, and bromadiolone were not degraded in the rumen but their bioavailabilities were high after oral administration; the kinetics of these compounds in sheep and other mammals are quite similar. These data suggest that the lack of susceptibility of ruminants to these anticoagulant rodenticides cannot be explained by either ruminal degradation or the specific toxicokinetics of these anticoagulants.

4-Hydroxycoumarins↗
PubMed1990

Effect of rotating monensin plus tylosin and lasalocid on performance, ruminal fermentation, and site and extent of digestion in feedlot cattle.

Two trials were conducted to evaluate the effects of ionophore rotation programs on performance and digestion by feedlot cattle. A 90% concentrate diet was fed with treatments of no ionophore (C), 33 mg lasalocid/kg diet daily (L), 29 mg monensin plus 11 mg tylosin/kg diet daily (MT), and daily (D) and weekly (W) rotation of L and MT. In Trial 1, feedlot performance of 200 crossbred steers (average initial BW 296 kg) was evaluated during a 133-d period. In Trial 2, four crossbred steers (average initial BW 376 kg) fitted with ruminal, duodenal and ileal cannulas were used in a 4 x 4 Latin square design to evaluate treatment effects (excluding W) on ruminal fermentation and site and extent of digestion. In Trial 1, daily rotation of L and MT improved (P less than .10) feed:gain ratio compared with other treatment groups, but daily feed intake did not differ (P greater than .10) among treatments. Daily gain was greater (P less than .10) for steers fed D than for those fed C or MT, but not different from that of steers fed L or W. Carcass measurements did not differ (P greater than .10) among treatments. In Trial 2, ruminal molar proportions of butyrate and valerate were decreased (P less than .07) by MT and D compared with C and L. Proportions of other VFA, ammonia concentrations and ruminal pH did not differ among treatments. Ionophore treatments did not affect site or extent of digestion of OM, starch or N; no differences among treatments were observed for efficiency of microbial protein synthesis. Although daily rotation of L and MT improved performance of growing-finishing feedlot steers, this improvement was not attributable to alterations in ruminal fermentation, or in site or extent of nutrient digestion.

Animals↗