Changes in the viscoelastic behaviour of the rumenal digesta mat of sheep fed on pasture or chaffed lucerne hay

We examined the changes in the physical properties of the digesta mat over a period of 24 h after cessation of feeding, in sheep that had been maintained on pasture or fed chaffed lucerne hay. The dry matter content of the digesta mat declined at similar rates in both dietary groups, although it was generally higher in sheep fed lucerne. Median particle size declined in the digesta mats of both dietary groups at similar rates in samples taken after 8 h, but median particle size was significantly greater in sheep fed chopped lucerne hay than in those fed grass. Thus, particles were not reduced to a common size suggesting that factors in addition to particle size governed the rate of breakdown of the rumen mat. The relationship between the elastic and loss moduli was of a consistent pattern in all samples taken from the rumen mat indicating that it behaved as a weak gel. The elastic and loss moduli of the digesta mats of sheep that had been fed pasture or chopped lucerne hay converged to similar values after 12 h and declined broadly at similar rates after this. The relationship between these two moduli and the dry matter content of the rumen mat were of similar curvilinear form for sheep on both diets. These findings suggest that the rate of breakdown of the rumen mat is more likely to be governed by its composite behaviour than by the size of the constituent particles.     

Effects of the feeding sequence of concentrate and forage and the feeding ratio of sake cake to grass hay on the characteristics and the entrapment effect of the ruminal mat in non‐lactating dairy cows

The effects of the feeding sequence of concentrate and forage, and of the feeding ratio of sake cake (SC) to grass hay (GH) on the ruminal mat characteristics and the entrapment effect for steamed flaked corn (SFC) by the mat were evaluated. Three non‐lactating, rumen‐cannulated Holstein cows were offered SC and GH using a ratio of 35:65 (SC35) or 65:35 (SC65). For SC35, SFC was fed at 1 h after or immediately before offering SC and GH, while for SC65, SFC was only fed at 1 h after. To estimate the degree of SFC entrapment in the ruminal mat, the location of SFC in the rumen and two types of mean retention time (MRT) were measured, that is, MRT for the SFC marker placed directly on the mat or taken orally. The ruminal mat was formed even when SC65 diet was consumed. The entrapment effect was not affected by the feeding sequence or the ratio. However, a more interesting finding is that the entrapment effect of the ruminal mat may not be as absolute as previously considered because of the large amount of SFC particles which precipitated at the bottom of the rumen 1 h after feeding.     

Effects of feeding ratio of beet pulp to alfalfa hay or grass hay on ruminal mat characteristics and chewing activity in Holstein dry cows

The influence of the feeding ratio of a non‐forage fiber source and hay on ruminal mat characteristics and chewing activity was evaluated in dairy dry cows. Cows were fed four different diets: the ratios of alfalfa hay (AH) to beet pulp (BP) were 8:2 (dry matter basis, A8B2) and 2:8 (A2B8), and those of grass hay (GH) to BP were 8:2 (G8B2) and 2:8 (G2B8). Total eating time was decreased with increasing BP content (P < 0.01). Total rumination time for AH was shorter than that for GH (P < 0.01), and it decreased with increasing BP content (P < 0.01). The ruminal mat was detected by using a penetration resistance test of the rumen digesta. Penetration resistance value (PRV) of ruminal mat was highest with the G8B2 diet and PRV decreased with increasing BP content (P < 0.05) and feeding AH (P < 0.05). Thickness of the ruminal mat was greater for increasing BP content (P < 0.05). Simple linear regression of ruminal mat PRV on total rumination time resulted in a high positive correlation (r = 0.744; P < 0.001; n = 16). The results demonstrated that increasing the PRV of the ruminal mat stimulated rumination activity and a ruminal mat could be formed, although it was soft even when cows were offered a large quantity of BP.     

Ruminal distribution of the cellulolytic bacterium Fibrobacter succinogenes in relation to its phylogenetic grouping

To investigate the ecological importance of the cellulolytic bacterium Fibrobacter succinogenes in fiber digestion, ruminal distribution of F. succinogenes was determined in relation to its phylogenetic grouping. Rumen digesta from wethers and steers fed orchardgrass hay, rice straw or fresh orchardgrass were employed as the materials for the analyses. Orchardgrass hay stem incubated in the rumen was also used. By using total DNA extracted from these materials, population sizes of total F. succinogenes and of four different phylogenetic groups of this species were quantitated through competitive polymerase chain reaction (PCR), and restriction fragment length polymorphism analysis of PCR products targeted the bacterial 16S rDNA. Rumen digesta and ruminally incubated hay stems had a reasonably high population size of F. succinogenes (×10^7?8/g) that was composed of strains belonging to the phylogenetic groups 1 and 3. The relative abundance of each group was different among the samples; group 1 dominated on the ruminally incubated hay stem and in the rumen of wethers fed fresh orchardgrass, while group 3 was major in the rumen of wethers and steers on hay diet. These results suggest that there could be phenotypic differences among the phylogenetic groups of F. succinogenes, and group 1 dominating on hay stem might contribute to rumen fiber digestion more than the other groups.     

Effect of a hay and a grain diet on the rate of hydrolysis of ochratoxin A in the rumen of sheep

The hydrolysis of ochratoxin A (OA) and the corresponding formation of its hydrolysis product, alpha ochratoxin (O alpha), by ruminal digesta and in the rumen of hay-fed and grain-fed sheep were compared. Ruminal contents from sheep fed diets with hay or with grain hydrolyzed OA in vitro; the majority of the activity was associated with the particulate fraction of the ruminal contents. The rate of hydrolysis of OA by ruminal fluid that was adjusted to different pH values was not influenced (P greater than .6) by the pH of the samples (pH was from 5.5 to 7.0). Ruminal fluid obtained from hay-fed animals (pH 7.0) was able to hydrolyze OA in vitro and to produce the hydrolyzed product, O alpha, at a much greater rate (fivefold) than ruminal fluid obtained from grain-fed animals (pH 5.5) (P less than .01). Ochratoxin A was administered intraruminally at a concentration of .5 mg/kg of BW to hay-fed and grain-fed sheep. The half-lives for disappearance of OA from the rumen of sheep fed grain (normal feed intake, rumen pH 5.7), fed grain at a low level (30% of normal feed intake, pH 6.5), and fed hay (pH 7.1) were 3.6, 1.3, and .6 h, respectively. The results suggest that OA is hydrolyzed much faster in the rumen of sheep fed hay than in sheep fed grain, presumably because of the different ruminal microbial population, which in turn influenced the rate of hydrolysis of OA.     

Effect of dietary protein level on nitrogen metabolism in the growing bovine: II. Diffusion into and utilization of endogenous urea nitrogen in the rumen

Six Angus heifer calves (234 kg) were assigned to either a high (HP; 126.1 g N/d) or low (LP; 66.5 g N/d) protein intake to evaluate ruminal criteria associated with movement of blood urea-N (BUN)-derived NH3-N from the rumen wall into interior ruminal digesta. Calves received 4.8 kg DM/d of diets containing 30% cottonseed hulls and 70% cornsoybean meal in equal portions at 4-h intervals. Following single i.v. injections of 15N-urea, ruminal fluid was collected serially for 4 h postinjection from digesta located adjacent to the rumen wall (wall-proximate digesta; WPD) and from the center of the rumen digesta mass after manual agitation (center mixed digesta; CMD). Mean ruminal NH3-N (RAN) concentrations were higher (P less than .05) for HP than for LP, but were not affected (P greater than .05) by digesta sampling site. Ruminal urease activity was higher (P less than .05) for LP than for HP and tended (P = .14) to be higher for WPD than for CMD. Area under the 15N enrichment curve (AUC) ratios between sampling sites (WPD/CMD x 100) for RAN were greater (P less than .05) for LP than for HP. However, AUC ratios for bacterial N were not affected (P greater than .05) by protein level. Whereas BUN-derived 15NH3 appeared to thoroughly equilibrate with RAN in interior ruminal digesta with HP, there appeared to be a declining enrichment gradient for RAN from the rumen wall to the interior ruminal digesta with LP. Data are interpreted to suggest that bacteria at or near the rumen wall may preferentially utilize some BUN-derived NH3-N entering through the rumen wall in calves fed LP diets.     

Effect of cereal straw and alfalfa hay diet on amino acid profile of gastrointestinal digesta in lactating dairy cows

This study was conducted to evaluate the effects of replacing alfalfa hay with rice straw (RS) or corn stover (CS) on amino acid (AA) profiles of gastrointestinal digesta in lactating cows. Eighteen lactating dairy cows were randomly assigned to one of three groups (n = 6) and fed identical concentrate and corn silage with different forages on dry matter basis: (i) 23% alfalfa hay and 7% Chinese wild rye hay (AH); (ii) 30% CS; and (iii) 30% RS. After the 14‐week feeding, a total of 18 cows were slaughtered to collect digesta from four representative organs, including rumen, omasum, duodenum and jejunum. The AA profiles of ruminal microbial fraction were similar among the treatments, except for greater Arg in cows fed RS than in cows fed AH or CS. Most of the analysed AA changed under different diets. Significant differences were found among the microbial fraction, rumen fluid and rumen digesta, with greater essential AA in digesta than in microbial fraction or rumen fluid and greater essential AA in microbial fraction than rumen fluid. Significant differences in individual AA profiles of digesta and relevant fluid were found across the four representative digestive tract parts, including rumen, omasum, duodenum and jejunum, showing much lower Leu proportion in CS and RS than in AH in duodenal fluid. In summary, ruminal microbes may prefer using essential AA, rather than non‐essential AA. The AA profile of ruminal microbes was constant except for Arg. The AA composition of digesta across the four digestive tracts changed dramatically, which indicated differences in the ability and efficiency of AA absorption. The lower duodenum absorbable Leu proportion in cows fed CS or RS indicated the shortage of Leu in CS or RS diets, which might also restrict the balanced AA absorption.     

Effects of time of day and monensin on the size distribution of particles in digestive tract sites of heifers fed corn silage

Effects of time of day and dietary monensin in the distribution of size of digesta particles in different digestive tract sites and their intersite relationships were examined in six heifers (290 kg BW) with ruminal, duodenal and ileal cannulas given ad libitum access to corn silage, with or without 100 mg monensin.head-1.d-1, in a two-period crossover design. Ingestive masticate and digesta of corn silage were collected via esophageal, ruminal or intestinal cannulas. The distribution of particulate matter retained on sieves with apertures larger than 20 microm was determined by wet-sieving. The cumulative distribution of particulate matter on a series of sieves was regressed on retaining sieve aperture to estimate the sieve aperture that would retain 50% weight of the particulate matter (median retaining aperture, MRA). The MRA of masticate was 6,494 microm. The MRA of digesta particles decreased (P less than .05) from ventral rumen (1,847 microm) to dorsal rumen (1,797 microm) to duodenum (346 microm), but increased to the rectum (359 microm). The MRA was lower (P = .044) for the monensin treatment only in feces. The MRA of particulate matter in the dorsal and ventral rumen, duodenum and rectum all changed (P less than .05) over 24 h. An inverse pattern between the MRA of ruminal and duodenal digesta occurred, presumably the result of a nycterohemeral pattern of eating and ruminating activity. Across sampling times, an inverse relationship existed between MRA of ventral rumen and duodenal digesta. This relationship suggests that a ruminal digesta raft composed of larger particles (immediately following major meals) is more effective than a raft of smaller particles (prior to such meals) in preventing flux of large particles to the duodenum.     

Influence of mass of ruminal contents on voluntary intake and digesta passage in steers fed a forage and a concentrate diet

To evaluate the influence of mass of ruminal contents on voluntary intake and ruminal function, five ruminally cannulated steers (550 kg) were fed an orchard grass hay diet ad libitum in a 5 x 5 Latin square experiment. The mass of ruminal contents was altered by adding varying weights of modified tennis balls to the rumen before the initiation of each 15-d experimental period. Treatments consisted of 50 balls with a specific gravity of 1.0, 1.1, 1.2, 1.3, or 1.4; the total weight of the balls was 7.45, 8.50, 9.25, 10.55, and 11.55 kg, respectively. Increasing the specific gravity of the balls added to the rumen decreased DMI and particle passage rate (P < 0.05) in a linear manner. A second experiment was conducted to evaluate the influence of mass of ruminal contents on voluntary intake and ruminal function of both forage and concentrate diets. Five ruminally cannulated steers (580 kg) were fed a 70% concentrate (DM basis) or an orchardgrass hay diet ad libitum in a 5 x 5 Latin square experiment. The mass of ruminal contents was altered as in the first experiment. Treatments consisted of 0 balls added to the rumen of steers fed concentrate diet (control), 75 balls with a specific gravity of 1.1 given to steers fed a concentrate diet, 75 balls with a specific gravity of 1.4 given to steers fed a concentrate diet, 75 balls with a specific gravity of 1.1 given to steers fed a hay diet, and 75 balls with a specific gravity of 1.4 given to steers fed hay diet. The addition of balls to the rumen of steers fed the concentrate diet decreased DMI (P < 0.05) compared with the 0-ball treatment, and increasing specific gravity of balls also decreased DMI (P < 0.01) for both concentrate and hay diets. Adding balls to the rumen of steers fed the concentrate diet decreased particle passage rate (P < 0.05), whereas increasing specific gravity of balls decreased particle passage rate for both concentrate and hay diet. The results of this study suggest that the density of ruminal digesta can have an influence on voluntary intake of both forage and concentrate diets.     

Influence of sampling site on concentrations and carbohydrate-degrading enzyme activities of protozoa and bacteria in the rumen.

Four ruminally cannulated cows were used to assess the distribution of the concentrations and carbohydrate-degrading enzyme activities of the liquid-associated protozoa (LAP) and solid-associated bacteria (SAB) in the rumen. The cows were fed diets (7 kg of DM/d) of 100% hay (Diet F) or 60% barley plus 40% hay (Diet C) in a 2 x 2 crossover experimental design. Samples of ruminal digesta were collected successively from the dorsal (DS), ventral (VS), and anterior (AS) sacs 1h before and 3 h after the morning feeding on two sampling days with a 2-d interval. Irrespective of diet and sampling time, the greatest proportion of entodiniomorphs, representing the main population of protozoa, was found in the DS (40% mean; P < .05). Low pH values observed in the DS (P < .05) indicated higher fermentative activity in this site than in the other parts of the rumen. Protozoa may contribute more than previously thought to the high digestive potential present at the top of the rumen that has classically been attributed to bacteria. The specific activity of plant cell wall polysaccharide-degrading enzymes in the LAP was correspondingly greater (P < .05) for DS than for VS or AS. For the two diets and sampling times, specific activity of fibrolytic enzymes in the SAB tended to be less in the upper than in the lower parts of the rumen, and less in the VS than in the AS. This tendency became significant (P < .05) for total fibrolytic enzyme activity. Differences in bacterial colonization of particles among the three sampling sites may explain such differences in fibrolytic activity of the SAB. Data suggest a spatio-temporal complementary action of the bacteria and protozoa in ruminal plant cell wall degradation, at least with the barley diet, for which the number of protozoa was highest.     

Comparison of rumen bacteria distribution in original rumen digesta,rumen liquid and solid fractions in lactating Holstein cows

Background: Original rumen digesta, rumen liquid and solid fractions have been frequently used to assess the rumen bacterial community. However, bacterial profiles in rumen original digesta, liquid and solid fractions vary from each other and need to be better established.Methods: To compare bacterial profiles in each fraction, samples of rumen digesta from six cows fed either a high fiber diet(HFD) or a high energy diet(HED) were collected via rumen fistulas. Rumen digesta was then squeezed through four layers of cheesecloth to separate liquid and solid fractions. The bacterial profiles of rumen original digesta, liquid and solid fractions were analyzed with High-throughput sequencing technique.Results: Rumen bacterial diversity was mainly affected by diet and individual cow(P 0.05) rather than rumen fraction. Bias distributed bacteria were observed in solid and liquid fractions of rumen content using Venn diagram and LEf Se analysis. Fifteen out of 16 detected biomarkers(using LEf Se analysis) were found in liquid fraction, and these 15 biomarkers contributed the most to the bacterial differences among rumen content fractions.Conclusions: Similar results were found when using samples of original rumen digesta, rumen liquid or solid fractions to assess diversity of rumen bacteria; however, more attention should be draw onto bias distributed bacteria in different ruminal fractions, especially when liquid fraction has been used as a representative sample for rumen bacterial study.     

Variation of bacterial communities and expression of Toll-like receptor genes in the rumen of steers differing in susceptibility to subacute ruminal acidosis

In order to determine differences in the ruminal bacterial community and host Toll-like receptor (TLR) gene expression of beef cattle with different susceptibility to acidosis, rumen papillae and content were collected from acidosis-susceptible (AS, n=3) and acidosis-resistant (AR, n=3) steers. The ruminal bacterial community was characterized using PCR-denaturing gradient gel electrophoresis (PCR-DGGE) and quantitative real time PCR (qRT-PCR) analysis. Global R analysis of bacterial profile similarity revealed that bacterial diversity was significantly different between AR and AS groups for both rumen content (P=0.001) and epithelial (P=0.002) communities. The copy number of total bacterial 16S rRNA genes in content of AS steers was 10-fold higher than that of AR steers, and the copy number of total 16S rRNA genes of epimural bacteria in AR steers was positively correlated with ruminal pH (r=0.59, P=0.04), and negatively correlated with total VFA concentration (r=-0.59, P=0.05). The expressions of host TLR2 and 4 genes were significantly higher in AR steers compared to those in AS steers. These findings enhance our understanding about the ruminal microbial ecology and host gene expression changes that may be useful in the prevention of ruminal acidosis.     

高谷物日粮对山羊小肠发酵、肠道结构和微生物菌群数量的影响研究

本试验旨在研究高谷物日粮对山羊小肠微生物发酵、上皮组织形态及微生物菌群数量的影响。采用随机区组实验设计,将12头山羊随机分为两组,即全干草组(只饲喂粗饲料)和高谷物组(75%精料和25%粗饲料混合饲喂),每组6头,试验期为6周,试验结束后屠宰取小肠内容物及组织样品用于相关分析。结果表明,1)与干草组相比,饲喂高谷物日粮显著提高了空肠内容物中总挥发性脂肪酸(P=0.015)、丙酸(P=0.008)、丁酸(P=0.004)、异丁酸浓度(P=0.035),降低了乳酸浓度(P=0.008),但对pH值、乙酸、戊酸、异戊酸浓度及LPS含量无显著性影响(P>0.05)。与干草组相比,饲喂高谷物日粮显著提高了回肠内容物的总挥发性脂肪酸(P=0.007)、丙酸(P=0.013)、丁酸(P=0.008)、戊酸(P<0.001)、乳酸浓度(P=0.008)以及脂多糖含量(P<0.001),降低了pH值(P=0.005),但对乙酸、异丁酸和异戊酸浓度无显著影响(P>0.05);2)与干草组相比,高谷物日粮组山羊的十二指肠、空肠和回肠的绒毛高度和隐窝深度均显著升高(P<0.001);空肠绒毛高度与隐窝深度的比值显著降低(P=0.024);电镜结果表明,高谷物组空肠和回肠紧密连接受到破坏;3)与干草组相比,高谷物日粮组山羊回肠黏膜中碱性磷酸酶活性显著提高(P<0.05),但对空肠黏膜中碱性磷酸酶活性无显著影响(P>0.05);4)Real-time PCR定量分析表明,与干草对照组相比,高谷物日粮山羊回肠拟杆菌门16S rRNA基因拷贝数显著降低(P=0.037),厚壁菌门与拟杆菌门细菌数量比值显著升高(P<0.001),但对厚壁菌门细菌基因拷贝数无显著影响(P>0.05);空肠中拟杆菌门基因拷贝数、厚壁菌门基因拷贝数以及厚壁菌门与拟杆菌门细菌数量比值无显著变化(P>0.05)。结果说明,饲喂高谷物日粮对回肠上皮组织形态及回肠微生物发酵具有显著影响,对其健康可能有不利影响。     

ORIGINAL ARTICLE: Ruminal and total tract phosphorus release from feedstuffs in cattle measured using the mobile nylon bag technique

Phosphorus (P) excretion in manure is a concern for dairy and beef producers. Excess P released into surface water runoff can lead to eutrophication and algal blooms in streams and lakes. One approach to reducing P excretion is to reduce dietary P. Data regarding P release from feedstuffs is limited and more precise formulations based on specific feed P release in the digestive tract may be one way to lower excreted P. In this experiment, the mobile nylon bag technique was used to determine the disappearance of P in corn silage, alfalfa hay, Coastal bermudagrass hay, and Tifton‐85 bermudagrass hay in steers after ruminal (24 h), ruminal + pepsin/HCl (rumen + PHCl), and ruminal + pepsin/HCl + intestinal (rumen + PHCl + I) incubation. Ruminal disappearance of P differed (p < 0.05) between feedstuffs and by site of incubation. Total tract (rumen + PHCl + I) P disappearance for corn silage, alfalfa hay, Coastal bermudagrass hay, and Tifton‐85 bermudagrass hay were 90.6%, 93.7%, 83.8% and 84.0% respectively. The range in P release (approximately 7%) indicates that considering P availability when balancing rations could have a measurable impact on subsequent P excretion from ruminants. More data concerning P availability as affected by other feed ingredients or plant species, maturity, and/or quality are needed to more accurately define P release from ruminant feeds.     

Research note: a method for studying local differences in ruminal fermentation in dairy cattle.

A method was developed for studying local differences in ruminal fermentation. The developed sampler consisted of an acrylic glass container (460 cm3) with an aperture for digesta sampling, which could be opened and closed by the scaled "T" rod. The scale was a reference for defined rumen layers: top, middle, 5 to 10 cm and 25 to 35 cm beneath the top of particles mat, respectively, and bottom 5 to 10 cm above the rumen floor. The repeatability of the method was proved in two rumen cannulated cows. Particle/fluid ratio, pH and sample amount were measured 2 to 2 1/2 h after morning feeding in four replicates each day (over 5 days), rumen layer and animal. No significant differences between replicates were observed. The coefficients of variation (CV) of the particle/fluid ratio varied between 8.7% and 13.6%. Top layer had higher CV than middle and bottom layer. CV of pH ranged between 0.59% and 1.27%. The developed method of sampling showed satisfactory repeatability for investigation of digesta properties and fermentation in different rumen layers.     

Effects of grass hay proportion in a corn silage‐based diet on rumen digesta kinetics and digestibility in dairy cows

In this study, we aimed to evaluate the effects of six levels of orchardgrass hay (GH) proportion (0%, 10%, 20%, 30%, 40% or 50% of dry matter) in finely chopped corn silage (CS)‐based diets on digesta kinetics of CS and GH in the rumen. Six non‐lactating, rumen‐cannulated Holstein cows were used in a 6 × 6 Latin square design. Ruminal digesta kinetics was measured by ruminal dosing of feed particle markers (dysprosium for CS, erbium for GH) followed by fecal sampling. The increase of GH proportion had a quadratic effect (P < 0.01) on total tract digestibility of neutral detergent fiber (NDF) and acid detergent fiber. The proportion of GH did not affect the particle size distribution of rumen digesta, total weight of dry matter or NDF in the rumen. The rates of large particle size reduction in the rumen for CS tended to increase linearly with increasing GH proportion (P = 0.077). A quadratic effect (P < 0.05) was found with increasing the GH proportion for the ruminal passage rate of small GH particles, but not for CS particles. The results suggested that associative effects between CS and GH could be generated on rumen digesta kinetics when cows were fed a CS‐based diet with an increased proportion of GH.     

Comparative digestion, rumen fermentation and kinetics of forage diets by steers and wethers

Four rumen fistulated wether and beef steers were used to evaluate differences in dry matter digestibility (DMD) between cattle and sheep. They were fed either perennial ryegrass or switchgrass hay at an ad libitum or restricted level for four experimental periods. Significant ruminant species X forage and ruminant species X level of intake (P less than .05) interactions were observed for digestible dry matter. The steers digested the switchgrass 7 percentage units greater than the wethers while ryegrass was digested equally. Digestibility differences between the steers and wethers were 6 percentage units at the ad libitum level of intake and 1 unit at the restricted level of intake. Crude protein digestibility tended to be greater (P less than .10) for sheep with a 7 unit difference for switchgrass and a 3 unit difference for ryegrass. The mean ruminal solids retention time of the digesta was approximately (P less than .01) 50% greater (26.0 vs 17.4 h) in cattle, with no difference in ruminal liquid dilution rate (LD) between animal species. Total ruminal volatile fatty acid concentration differed (P less than .01) with level of intake; however, no influence due to intake on the molar proportion of acetate (P greater than .10) or propionate (P greater than .10) was evident in spite of a difference (P less than .01) in LD. Rumen pH (P less than .05) and osmolality (P less than .01) were affected by both level of intake and forage, with the ryegrass and high level of intake decreasing pH and increasing osmolality.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of slaframine on ruminant digestive function: liquid turnover rate and fermentation patterns in sheep and cattle

Two trials were initiated to determine if slaframine (SF) can be used to alter fluid digesta flow and fermentation patterns in the rumen. In trial 1, a preliminary experiment, four Dorset X Barbados Black-belly ruminal-cannulated wethers (avg weight 41.6 8.7 kg) given ad libitum access to a pelleted concentrate/hay diet were injected intramuscularly with 0, 12, 24 or 48 micrograms SF/kg body weight (BW) in a 4 X 4 Latin-square design. Ruminal fluid dilution rate was determined using a single intraruminal infusion of polyethylene glycol (7 g), followed by seven hourly ruminal fluid samples. The administration of 48 micrograms SF/kg BW increased (P less than .10) ruminal volume and outflow by 27 and 25%, respectively, compared with controls. In trial 2, two Hereford and two Angus ruminal cannulated steers (avg weight 568 +/- 93 kg) were injected with 0, 6, 12 or 24 micrograms SF/kg BW at 8-h intervals over a 24-h period in a 4 X 4 Latin-square design. Steers were fed a concentrate diet at twice maintenance in 24 equal portions daily. Ruminal fluid dilution was measured using a single intraruminal infusion of cobalt-ethylenediamine tetraacetic acid (20 g) administered 9 h after the initial SF injection. Ruminal fluid was collected each hour during 8 to 24 h after the initial SF injection and analyzed for pH, osmolality and volatile fatty acids (VFA).(ABSTRACT TRUNCATED AT 250 WORDS)     

Characteristics of water soluble markers for measuring rumen liquid volume and dilution rate

The utility of five water soluble markers (WSM) for measuring rumen liquid volume and dilution rate was examined in nine in vitro and in vivo experiments. The WSM tested included polyethylene glycol (PEG) and ethylenediaminetetraacetic acid (EDTA) complexes of Cr, Co, Fe and Yb. Feedstuffs incubated in vitro absorbed variable amounts of distilled H2O and autoclaved rumen fluid with roughages imbibing greater (P less than .05) amounts of fluid than feeds higher in starch. Liquid present in the feeds tested reached equilibrium with added WSM in about 20 min. Corn grain in the whole or ground form appeared to exclude 4 to 16% of the WSM while imbibing water. Cottonseed hulls reduced detectable PEG due to some water soluble substance present. The WSM tested did not produce different estimates of dilution rate (P = .65) and volume (p = .29) of steers fed either 80% whole shelled corn, 90% chopped alfalfa hay or 90% prairie hay diets. Ruminal liquid measured by hand removal was about 4% greater than volume estimated by WSM. The percentage of total ruminal water associated with solids varied from 27 to 86% among steers. Compared with steers fed an 80% whole shelled corn diet, steers fed equal dry matter from alfalfa hay had 15% more dry matter in the rumen, 21% larger liquid volume, 80% greater ruminal liquid dilution rate and 116% greater rate of liquid flow from the rumen. Dilution rate of ruminal liquid during feeding was examined with six steers fed a 90% chopped alfalfa hay diet once daily at 1.8% of body weight. Dilution rate estimated with Co EDTA increased by 166% during the 4 h after feeding. The WSM did not influence (P greater than .10) digestibility of dry matter with steers fed a 90% roughage or a 90% concentrate ration. Recovery of WSM administered via a rumen cannula ranged from 96.8 to 99.7% in feces and 1.2 to 3.4% in urine. All WSM examined appeared suitable for estimating rumen liquid volume and dilution rate for diets not containing cottonseed hulls.     

Characterization of bovine ruminal epithelial bacterial communities using 16S rRNA sequencing, PCR-DGGE, and qRT-PCR analysis

Currently, knowledge regarding the ecology and function of bacteria attached to the epithelial tissue of the rumen wall is limited. In this study, the diversity of the bacterial community attached to the rumen epithelial tissue was compared to the rumen content bacterial community using 16S rRNA gene sequencing, PCR-DGGE, and qRT-PCR analysis. Sequence analysis of 2785 randomly selected clones from six 16S rDNA (～1.4kb) libraries showed that the community structures of three rumen content libraries clustered together and were separated from the rumen tissue libraries. The diversity index of each library revealed that ruminal content bacterial communities (4.12/4.42/4.88) were higher than ruminal tissue communities (2.90/2.73/3.23), based on 97% similarity. The phylum Firmicutes was predominant in the ruminal tissue communities, while the phylum Bacteroidetes was predominant in the ruminal content communities. The phyla Fibrobacteres, Planctomycetes, and Verrucomicrobia were only detected in the ruminal content communities. PCR-DGGE analysis of the bacterial profiles of the rumen content and ruminal epithelial tissue samples from 22 steers further confirmed that there is a distinct bacterial community that inhibits the rumen epithelium. The distinctive epimural bacterial communities suggest that Firmicutes, together with other epithelial-specific species, may have additional functions other than food digestion.