Effects of condensed tannins in wrapped silage bales of sainfoin (Onobrychis viciifolia) on in vivo and in situ digestion in sheep

The objective of this study was to characterize the condensed tannins (CTs) in wrapped silage bales of sainfoin (Onobrychis viciifolia) and examine their potential action on in vivo and in situ digestive characteristics in sheep. Silage was made from sainfoin, cut at two phenological stages. The first phenological stage, at which silage was made, was from the first vegetation cycle at the end of flowering and the second stage silage was made from regrowth, 5 weeks after the first cut, but before flowering. The silages made from the two phenological stages were fed to 12 rumen-fistulated sheep in a crossover design. Of the 12 sheep, six received polyethylene glycol (PEG) to bind with and remove the effects of CT, whereas the other six were dosed with water. Organic matter digestibility, total-tract N digestibility and N (N) balance were measured over 6 days. Kinetic studies were performed on total N, ammonia N (NH3-N) and volatile fatty acids (VFAs) in rumen fluid before and 1.5, 3 and 6 h after feeding. The kinetics of degradation of dry matter and N from Dacron bags suspended in the rumen were also determined. Biological activity of CT (protein-binding capacity) and CT concentration were greater for the silage made from sainfoin at the early flowering stage. Total-tract N digestibility was increased by the addition of PEG (P < 0.001) to the sainfoin silage before flowering (P < 0.001). CTs decreased N excretion in urine (P < 0.05) and increased faecal N excretion (P < 0.001), but had no effect on body N retention, which is beneficial for the animal. Ruminal N degradability was smaller in the presence of active CT (P < 0.001) at both phenological stages; however, soluble N (P = 0.2060) and NH3-N (P = 0.5225) concentrations in rumen fluid remained unchanged. The results of this experiment indicate that CT in the sainfoin retain their ability to affect the nutritive value of preserved forage legumes.     

Bacterial communities in the rumen and feces of lactating Holstein dairy cows are not affected when fed reduced-fat dried distillers’ grains with solubles

Reduced-fat dried distillers’ grains with solubles (RF-DDGSs) are co-products of ethanol production and contain less fat than traditional distillers’ grains. The fat in corn is ~91% unsaturated, and it is toxic to rumen microorganisms so it could influence the composition of the rumen microbiome. It has been demonstrated that RF-DDGS is a suitable ration ingredient to support the high-producing dairy cow, and this feedstuff is a promising alternative protein source for lactating dairy cows. The current study aims to better understand the effect of RF-DDGS on the rumen and fecal bacterial composition in lactating dairy cows. Thirty-six multiparous (two or three), mid-lactation Holstein cows (BW = 680 ± 11 kg; 106 ± 27 DIM) were randomly assigned to two groups which were fed a control diet made up of corn, corn silage, and alfalfa hay supplemented with expeller soybean meal or with added RF-DDGS (20% of the DM) containing approximately 6.0% fat. Whole rumen contents (rumen fluid and digesta; esophageal tubing method) and feces (free-catch method) were collected on day 35 of the experimental period, after the 14-d acclimation period. Rumen contents and feces from each cow were used for DNA extraction. The bacterial community composition in rumen and fecal samples was assessed via the 16S rRNA gene by using the Illumina MiSeq sequencing platform. Bacteroidetes, Actinobacteria, and Firmicutes were the most abundant phyla in rumen contents. The fecal microbiota was dominated by the phyla Firmicutes and Bacteroidetes, as well as Actinobacteria and Chloroflexi. RF-DGGS increased bacterial richness, evenness, and Shannon diversity in both rumen and fecal samples and was associated with several taxa that had different abundance in treatment versus control comparisons. The RF-DGGS, however, did not significantly alter the bacterial community in the rumen or feces. In general, these findings demonstrated that dietary inclusion of RF-DDGS did not impose any serious short-term (within 30 days) health or production consequences, as would be expected. With this study, we present further evidence that inclusion of 20% (DM basis) RF-DDGS in the diet of lactating dairy cows can be done without consequence on the microbiome of the rumen.     

Effect of feeding dehydrated and ensiled tanniferous sainfoin (Onobrychis viciifolia) on nitrogen and mineral digestion and metabolism of lambs

The effects of tanniferous sainfoin on digestion and metabolism have been investigated in 12 lambs in an incomplete cross-over design (n = 6). Effects of condensed tannins (CT) were evaluated by comparing dehydrated and ensiled sainfoin treated with and without polyethylene glycol (PEG). Dehydrated and ensiled grass-clover mixtures served as controls. The lambs were fed the treatment diets, including a mineral supplement, for 21 d. During the last 7 d excreta, rumen fluid and blood were sampled. The CT of sainfoin decreased rumen fluid ammonia concentration (p < 0.001) and increased the plasma concentration mainly of essential amino acids (p < 0.001). Body retention of phosphorus, calcium and magnesium was lower with sainfoin compared to PEG-treated sainfoin (p < 0.05). Sainfoin without PEG resulted in lower digestibilities of organic matter and neutral detergent fibre than sainfoin with PEG and the grass-clover mixture (p < 0.001). Ensiling of sainfoin led to the lowest N-retention. In conclusion, the reduction in ruminal ammonia and urine-N losses by sainfoin CT did not improve N-retention.     

Lotus corniculatus condensed tannins decrease in vivo populations of proteolytic bacteria and affect nitrogen metabolism in the rumen of sheep

Condensed tannins in forage legumes improve the nutrition of sheep by reducing ruminal degradation of plant protein and increasing crude protein flow to the intestine. However, the effects of condensed tannins in forage legumes on rumen bacterial populations in vivo are poorly understood. The aim of this study was to investigate the specific effects of condensed tannins from Lotus corniculatus on four proteolytic rumen bacteria in sheep during and after transition from a ryegrass (Lolium perenne)-white clover (Trifolium repens) diet (i.e., low condensed tannins) to a Lotus corniculatus diet (i.e., higher condensed tannins). The bacterial populations were quantified using a competitive polymerase chain reaction. Lotus corniculatus was fed with or without ruminal infusions of polyethylene glycol (PEG), which binds to and inactivates condensed tannins, enabling the effect of condensed tannins on bacterial populations to be examined. When sheep fed on ryegrass-white clover, populations of Clostridium proteoclasticum B316T, Butyrivibrio fibrisolvens C211a, Eubacterium sp. C12b, and Streptococcus bovis B315 were 1.5 x 10(8), 1.1 x 10(6), 4.6 x 10(8), and 7.1 x 10(6) mL(-1), respectively. When the diet was changed to Lotus corniculatus, the average populations (after 8-120 h) of C. proteoclasticum, B. fibrisolvens, Eubacterium sp., and S. bovis decreased (P < 0.001) to 2.4 x 10(7), 1.1 x 10(5), 1.1 x 10(8), and 2.5 x 10(5) mL(-1), respectively. When PEG was infused into the rumen of sheep fed Lotus corniculatus, the populations of C. proteoclasticum, B. fibrisolvens, Eubacterium sp., and S. bovis were higher (P < 0.01-0.001) than in sheep fed Lotus corniculatus without the PEG infusion, with average populations (after 8-120 h) of 4.9 x 10(7), 3.8 x 10(5), 1.9 x 10(8), and 1.0 x 10(6), respectively. Sheep fed the Lotus corniculatus diet had lower rumen proteinase activity, ammonia, and soluble nitrogen (P < 0.05-0.001) than sheep that were fed Lotus corniculatus plus PEG. The Lotus corniculatus diet reduced rumen nitrogen digestibility (P < 0.05) and ammonia pool size and increased the flow of undegraded feed nitrogen to the abomasum. The nitrogen intake, rumen non-ammonia nitrogen pool size, rumen microbial non-ammonia nitrogen pool size, and abomasal microbial non-ammonia nitrogen fluxes were similar both in sheep fed only Lotus corniculatus and in sheep fed Lotus corniculatus plus PEG, but nonmicrobial non-ammonia nitrogen flux to the abomasum was higher (P < 0.01) for the sheep fed only Lotus corniculatus. Although condensed tannins in Lotus corniculatus reduced the populations of some proteolytic bacteria, total ruminal microbial protein and microbial protein outflow to the abomasum were unchanged, suggesting a species-specific effect of condensed tannins on bacteria in the rumen.     

Potential To Reduce Escherichia coli Shedding in Cattle Feces by Using Sainfoin (Onobrychis viciifolia) Forage, Tested In Vitro and In Vivo

There is a growing concern about the presence of pathogens in cattle manure and its implications on human and environmental health. The phytochemical-rich forage sainfoin (Onobrychis viciifolia) and purified phenolics (trans-cinnamic acid, p-coumaric acid, and ferulic acid) were evaluated for their ability to reduce the viability of pathogenic Escherichia coli strains, including E. coli O157:H7. MICs were determined using purified phenolics and acetone extracts of sainfoin and alfalfa (Medicago sativa), a non-tannin-containing legume. Ground sainfoin or pure phenolics were mixed with fresh cattle feces and inoculated with a ciprofloxacin-resistant strain of E. coli, O157:H7, to assess its viability at −20°C, 5°C, or 37°C over 14 days. Forty steers were fed either a sainfoin (hay or silage) or alfalfa (hay or silage) diet over a 9-week period. In the in vitro study, the MICs for coumaric (1.2 mg/ml) and cinnamic (1.4 mg/ml) acids were 10- to 20-fold lower than the MICs for sainfoin and alfalfa extracts. In the inoculated feces, the −20°C treatment had death rates which were at least twice as high as those of the 5°C treatment, irrespective of the additive used. Sainfoin was less effective than coumaric acid in reducing E. coli O157:H7 Cip^r in the inoculated feces. During the animal trial, fecal E. coli numbers declined marginally in the presence of sainfoin (silage and hay) and alfalfa silage but not in the presence of hay, indicating the presence of other phenolics in alfalfa. In conclusion, phenolic-containing forages can be used as a means of minimally reducing E. coli shedding in cattle without affecting animal production.     

Solubility and flow of calcium,magnesium and phosphorus in the digestive tract of sheep given maize or alfalfa silages

Four sheep, each fitted with a rumen cannula and a re-entrant cannula in the proximal duodenum and terminal ileum, were used for collection of rumen, duodenal and ileal digesta. Four normal sheep were used for collection of faeces. They were given maize silage, maize silage ensiled with urea, wilted alfalfa silage or formic acid-treated alfalfa silage. Although the solubility of the calcium (Ca) in the duodenal digesta was higher with sheep on maize silages than with those on alfalfa silages, sheep fed on maize silage showed negative apparent absorption (?32.7 and ?16.5%) owing to the low concentration of Ca in whole-plant maize. The apparent absorption of magnesium (Mg) was 40.6–42.3% for maize silages and 24.2–26.8% for alfalfa silages. The differences between silages with respect to apparent absorption of phosphorus (P) were not significant and ranged between 1.5 and 10.9%. There were no appreciable differences in soluble proportions, flow and apparent absorption of Ca, Mg and P between the two maize silages or the two alfalfa silages. There was a net absorption of Ca and Mg and a net secretion of P in the stomach of animals on all silages.     

Rumen microbial changes in cattle fed diets with or without salinomycin.

Four rumen-fistulated steers, randomly assigned to two groups (control and salinomycin fed) were used to monitor the changes in rumen microbial populations and volatile fatty acids (VFA) concentrations associated with feeding salinomycin (0.22 mg X kg-1 X day-1). Steers were adapted to an alfalfa hay and grain (80:20) diet before supplementing the diet with salinomycin, and then the diet was changed to 50:50 and 20:80 ratios of alfalfa hay to grain at 2-week intervals. Rumen samples for total and selective enumeration of anaerobic bacteria. VFA analysis, and enumeration of protozoa were collected during the 80:20 alfalfa hay-to-grain diet before salinomycin feeding, and during the 80:20, 50:50, and 20:80 hay-to-grain diets with salinomycin. At each sampling period, rumen samples were collected at 3 h after feeding on three consecutive days. Salinomycin feeding had no effect on rumen pH and total VFA concentration. The acetate-to-propionate ratio was significantly lower in salinomycin-fed steers than in the control. The molar proportion of butyrate increased in both control and salinomycin-fed steers. Total anaerobic bacterial counts were lower in salinomycin-fed steers than in the control steers after 8 weeks of salinomycin feeding. Salinomycin-resistant bacteria increased from 7.6 to 15.6% in salinomycin-fed steers but remained unchanged in control steers. Salinomycin had no effect on cellulolytic and lactate-utilizing bacteria, but the proportion of amylolytic bacteria was higher in salinomycin-fed steers than in control steers. The total number of protozoa decreased initially in salinomycin-fed steers. The initial reduction was due to reduced numbers of Entodinium species. Holotrichs were unaffected by salinomycin feeding.     

Rumen microbial changes in cattle fed diets with or without salinomycin

Four rumen-fistulated steers, randomly assigned to two groups (control and salinomycin fed) were used to monitor the changes in rumen microbial populations and volatile fatty acids (VFA) concentrations associated with feeding salinomycin (0.22 mg X kg-1 X day-1). Steers were adapted to an alfalfa hay and grain (80:20) diet before supplementing the diet with salinomycin, and then the diet was changed to 50:50 and 20:80 ratios of alfalfa hay to grain at 2-week intervals. Rumen samples for total and selective enumeration of anaerobic bacteria. VFA analysis, and enumeration of protozoa were collected during the 80:20 alfalfa hay-to-grain diet before salinomycin feeding, and during the 80:20, 50:50, and 20:80 hay-to-grain diets with salinomycin. At each sampling period, rumen samples were collected at 3 h after feeding on three consecutive days. Salinomycin feeding had no effect on rumen pH and total VFA concentration. The acetate-to-propionate ratio was significantly lower in salinomycin-fed steers than in the control. The molar proportion of butyrate increased in both control and salinomycin-fed steers. Total anaerobic bacterial counts were lower in salinomycin-fed steers than in the control steers after 8 weeks of salinomycin feeding. Salinomycin-resistant bacteria increased from 7.6 to 15.6% in salinomycin-fed steers but remained unchanged in control steers. Salinomycin had no effect on cellulolytic and lactate-utilizing bacteria, but the proportion of amylolytic bacteria was higher in salinomycin-fed steers than in control steers. The total number of protozoa decreased initially in salinomycin-fed steers. The initial reduction was due to reduced numbers of Entodinium species. Holotrichs were unaffected by salinomycin feeding.     

In vivo and in situ measurements of the digestive characteristics of sainfoin in comparison with lucerne fed to sheep as fresh forages at two growth stages and as hay

In vivo and in situ digestive characteristics of sainfoin (Onobrychis viciifolia L., a tannin-rich forage) and lucerne (Medicago sativa L., a tannin-free forage) were compared to evaluate the effects of condensed tannins (CT) and growth stage (vegetative v. early flowering) in experiment 1. In experiment 2, the hays of the two forages, harvested at early flowering, were compared. Ingestibility, organic matter digestibility (OMD) and nitrogen (N) retention were measured in sheep fed sainfoin and lucerne fresh forages and hays. The loss of dry matter (DM) and N from polyester bags suspended in the rumen, abomasum and small intestine was also measured using rumen fistulated sheep and other intestine fistulated sheep. Nitrogen content was lower in sainfoin than in lucerne. Content of CT in sainfoin decreased with growth stage (3.5 to 2.5 g CT/kg DM) and was lower for sainfoin hay (0.6 g CT/kg DM). Ingestibility and OMD did not differ between fresh-fed forage species. Total N tract digestibility in vivo was much lower for sainfoin than for lucerne fresh forages (mean value 0.540 v. 0.721, P < 0.001) and for sainfoin hay than lucerne hay (0.464 v. 0.683, P < 0.001). In both species, N digestibility was not altered by growth stage. The rumen degradation of N was lower in sainfoin than in lucerne, resulting in a lower proportion of N intake excreted in urine. The intestinal digestibility of sainfoin was also lower than that of lucerne, resulting in a higher N excretion in faeces. Hence the efficiency of N utilisation by sheep (ENr) was similar (mean value 0.205 and 0.199 g N retained/g N intake for fresh sainfoin and lucerne, respectively). The coefficient of N retention by the animal was higher for sainfoin at the vegetative stage than for all the other forages. Nitrogen degradability in the rumen determined by the nylon bag technique (DegN) was lower for sainfoin than for lucerne when forages were studied both fresh (mean value 0.608 and 0.818, respectively) and as hays (0.631 and 0.767). The efficiency of forage N digestion (ENd) was higher for sainfoin at the vegetative stage. Compared with lucerne, sainfoin greatly increased the in situ estimate of forage N escaping the rumen but decreased its intestinal digestibility.     

Effect of a fibrolytic enzyme preparation from Trichoderma longibrachiatum on the rumen microbial population of dairy cows

The effects of supplementing a dairy cow diet with incremental levels of a fibrolytic enzyme preparation (preparation B) from Trichoderma longibrachiatum on the rumen microbial population were investigated. Two cows fitted with rumen cannulae were each fed a diet containing barley-based concentrate (52%), maize silage (29%), and chopped alfalfa hay (19%), supplemented with 0, 1, 2, 5, or 10 L of preparation B per tonne of dry matter (DM). Preparation B stimulated numbers of total viable bacteria in a quadratic manner (P < 0.05), to approximately 230, 330, 390, and 250% at 1, 2, 5, and 10 L x t(-1) DM, respectively. Preparation B increased the numbers of cellobiose-utilizing (P < 0.01), xylanolytic (P < 0.05), and amylolytic bacteria (P < 0.05), but had no effect (P > 0.05) on numbers of cellulolytic bacteria. However, when bacterial numbers enumerated on each substrate were expressed as a proportion of total viable bacterial numbers, only cellobiose utilizers were stimulated, and this stimulation was limited to the 1 L x t(-1) DM level of preparation B (P < 0.05). The results of this study demonstrate that the inclusion of an exogenous fibrolytic enzyme preparation in dairy cow diets increased the numbers of rumen bacteria that utilize hemicelluloses and secondary products of cellulose digestion.     

Effects of dried distillers' grain on fecal prevalence and growth of Escherichia coli O157 in batch culture fermentations from cattle

Distillers' grains (DG), a by-product of ethanol production, are fed to cattle. Associations between Escherichia coli O157 prevalence and feeding of DG were investigated in feedlot cattle (n = 379) given one of three diets: steam-flaked corn (SFC) and 15% corn silage with 0 or 25% dried distillers' grains (DDG) or SFC with 5% corn silage and 25% DDG. Ten fecal samples were collected from each pen weekly for 12 weeks to isolate E. coli O157. Cattle fed 25% DDG with 5 or 15% silage had a higher (P = 0.01) prevalence of E. coli O157 than cattle fed a diet without DDG. Batch culture ruminal or fecal microbial fermentations were conducted to evaluate the effect of DDG on E. coli O157 growth. The first study utilized microbial inocula from steers fed SFC or dry-rolled corn with 0 or 25% DDG and included their diet as the substrate. Ruminal microbial fermentations from steers fed DDG had higher E. coli O157 contents than ruminal microbial fermentations from steers fed no DDG (P < 0.05) when no substrate was included. Fecal fermentations showed no DDG effect on E. coli O157 growth. In the second study with DDG as a substrate, ruminal fermentations with 0.5 g DDG had higher (P < 0.01) E. coli O157 concentrations at 24 h than ruminal fermentations with 0, 1, or 2 g DDG. In fecal fermentations, 2 g DDG resulted in a higher concentration (P < 0.05) at 24 h than 0, 0.5, or 1 g DDG. The results indicate that there is a positive association between DDG and E. coli O157 in cattle, and the findings should have important ramifications for food safety.     

乙醇对瘤胃液接种稻秸的体外发酵产物及细菌群落结构的影响

旨在研究乙醇对山羊瘤胃液与水稻秸秆厌氧共培养的影响。利用频繁传代的体外发酵技术和高通量测序方法,分析了短链脂肪酸(SCFA)产量和细菌群落的变化。结果表明,经体外培养传代8次的稻秸发酵液的总短链脂肪酸产量显著高于瘤胃液(P<0.01);与未添加乙醇的稻秸发酵液相比,添加乙醇显著提高了乙酸、戊酸和己酸的比例,降低了丙酸和丁酸的比例(P<0.01),总SCFA产量及异丁酸和异戊酸比例无显著差异。与瘤胃液相比,稻秸发酵液的拟杆菌门(Bacteroidetes)相对丰度下降,厚壁菌门(Firmicutes)相对丰度升高(P<0.05),且添加乙醇显著提高了厚壁菌门和放线菌门(Actinobacteria)的相对丰度(P<0.05);添加乙醇使双歧杆菌属(Bifidobacterium)、未定性的毛螺菌属(unidentified Lachnospiraceae)、产琥珀酸菌属(Succiniclasticum)、脱硫弧菌属(Desulfovibrio)和未定性的梭菌属(unidentified Clostridiales)的相对丰度显著升高(P<0.05)。乙醇使稻秸发酵液的显著性差异物种(Biomarker)增加;稻秸发酵液与瘤胃液亲缘关系较近,而添加乙醇显著改变了细菌区系;短链脂肪酸比例在稻秸发酵液细菌群落多样性中具有重要作用。研究表明,体外频繁传代和添加乙醇可以提高稻秸发酵液的乙酸、戊酸和己酸产量,乙醇改变了稻秸发酵液的细菌群落结构。     

Microbial inoculant effects on silage and in vitro ruminal fermentation, and microbial biomass estimation for alfalfa, bmr corn, and corn silages

Whole crop third cut alfalfa, brown mid-rib (bmr) corn, and corn were chopped and inoculated with one of four microbial inoculants used. Uninoculated silage was the control treatment. Each crop was ensiled in four mini-silos (1 L glass jars) per treatment. All silos were fermented for 60 days at room temperature (22 °C), and then they were opened and analyzed for fermentation products, fiber constituents and N fractions. A fraction of wet silage was ground with a blender for 30 s. In vitro gas production was measured in 160 ml sealed serum vials at 3, 6, 9, 24, and 48 h using the wet ground silage. At 9 and 48 h, rumen fluid was analyzed for volatile fatty acids (VFA) and microbial biomass yield (MBY). In all the three crops, the four inoculants produced only minor changes in pH and fermentation products during ensiling. Of the variables measured, soluble nonprotein N fractions were the characteristics most often affected by some inoculants. At 9 h incubation, in vitro gas production and VFA did not differ between control and inoculated silages, but MBY did. Among crops, alfalfa and corn silages had higher MBY than did bmr corn silage. Among inoculants, three of the four inoculated silages produced more MBY than did control. At 48 h, alfalfa silage produced higher MBY than did corn or bmr silage, and two of the inoculated silages had more MBY than did the control. There was no inoculant by crop interaction. Results suggest that some silage inoculants are capable of altering rumen fermentation, even in cases where effects on silage fermentation are small, and that this effect may be linked to better preservation of crop protein during ensiling.     

Effect of partially replacing a barley‐based concentrate with flaxseed‐based products on the rumen bacterial population of lactating Holstein dairy cows

Aims

The effects of partial replacement of a barley‐based concentrate with flaxseed‐based products on the rumen bacterial population of lactating Holstein dairy cows were evaluated.

Methods and Results

Treatments fed were CONT, a normal diet that included barley silage, alfalfa hay and a barley‐based concentrate that contained no flaxseed or faba beans; FLAX, inclusion of a nonextruded flaxseed‐based product containing 55·0% flaxseed, 37·8% field peas and 6·9% alfalfa; EXT, similar to FLAX, but the product was extruded and EXTT, similar to FLAX, but product was extruded and field peas were replaced by high‐tannin faba beans. The rumen bacterial population was evaluated by utilizing 16S rRNA gene sequencing. Most abundant phyla, families and genera were unaffected. However, some taxa were affected; for example, unsaturated fatty acid content was negatively correlated with Clostridiaceae, and tannin content was negatively correlated with BS11 and Paraprevotellaceae.

Conclusions

Predominant rumen bacterial taxa were not affected, but the abundance of some taxa found in lower proportions shifted, possibly due to sensitivity to unsaturated fatty acids or tannins.

Significance and Impact of the Study

Flaxseed‐based products were effective for partially replacing barley‐based concentrate in rations of lactating dairy cows. No negative effects of these products were observed on the abundance of predominant rumen bacterial taxa, with only minor shifts in less abundant bacteria.     

High-Throughput DNA Sequencing of the Ruminal Bacteria from Moose (Alces alces) in Vermont,Alaska, and Norway

In the present study, the rumen bacteria of moose (Alces alces) from three distinct geographic locations were investigated. Moose are large, browsing ruminants in the deer family, which subsist on fibrous, woody browse, and aquatic plants. Subspecies exist which are distinguished by differing body and antler size, and these are somewhat geographically isolated. Seventeen rumen samples were collected from moose in Vermont, Alaska, and Norway, and bacterial 16S ribosomal RNA genes were sequenced using Roche 454 pyrosequencing with titanium chemistry. Overall, 109,643 sequences were generated from the 17 individual samples, revealing 33,622 unique sequences. Members of the phylum Bacteroidetes were dominant in samples from Alaska and Norway, but representatives of the phylum Firmicutes were dominant in samples from Vermont. Within the phylum Bacteroidetes, Prevotellaceae was the dominant family in all three sample locations, most of which belonged to the genus Prevotella. Within the phylum Firmicutes, the family Lachnospiraceae was the most prevalent in all three sample locations. The data set supporting the results of this article is available in the Sequence Read Archive (SRA), available through NCBI [study accession number SRP022590]. Samples clustered by geographic location and by weight and were heterogenous based on gender, location, and weight class (p?<?0.05). Location was a stronger factor in determining the core microbiome than either age or weight, but gender did not appear to be a strong factor. There were no shared operational taxonomic units across all 17 samples, which indicates that these moose may have been isolated long enough to preclude a core microbiome among moose. Other potential factors discussed include differences in climate, food quality and availability, gender, and life cycle.     

Effect of rumen ciliates on the digestion of different carbohydrates in sheep. I.--Utilization of cell wall carbohydrates (cellulose and hemicellulose) and of starch

Two diets rich in cell-wall carbohydrates or starch were given to 10 rumen-fistulated sheep; two sheep were defaunated and the others were inoculated with Polyplastron multivesiculatum (P) or Entodinium sp. (E), or both (P + E), or with conventional fauna. Ciliate biomass was greater when the animals were fed a high starch diet than when the diet was rich in cell-wall carbohydrates (table 2). With both diets, the Entodinium genus in the mixed fauna sampling predominated. We showed that Polyplastron was directly involved in cell-wall carbohydrate breakdown, while Entodinium capacity to digest cellulose remained low. We noted that with a diet rich in cellulose and hemicellulose, bacterial cellulolytic activity was improved by the presence of ciliates in the rumen but was decreased with the "starch" diet (table 3). The greater VFA concentration observed in the faunated animals expressed ciliate effect on the fermentations as well as activation of bacterial metabolism. With a high starch diet, the Entodinium sp. ciliates may have a buffering effect on the pH values in the rumen by limiting bacterial fermentation after food intake and by prolonging starch digestion during the day (table 4). The composition of the VFA mixture was modified by ciliate inoculation. The molar proportion of butyric acid always increased, while that of acetic and propionic acids evolved differently according to the diets and the ciliates (table 4). The higher ammonia concentration in the rumen liquor observed in faunated animals (table 4) could be explained either by the breakdown of both feed and bacterial proteins ingested by ciliates or by a lower ammonia nitrogen incorporation by fewer bacteria. Statistical analyses were used to explain the specific effect of P and E and also the interactions between them and between each of them and the diets.     

Fenugreek (Trigonella foenum-graecum L.) as an alternative forage for dairy cows

Fenugreek is a novel forage crop in Canada that is generating interest as an alternative to alfalfa for dairy cows. To evaluate the value of fenugreek haylage relative to alfalfa haylage, six, second lactation Holstein cows (56 ± 8 days in milk), which were fitted with rumen cannulas (10 cm i.d., Bar Diamond Inc., Parma, ID, USA) were used in a replicated three × three Latin square design with 18-day periods. Diets consisting of 400 g/kg haylage, 100 g/kg barley silage and 500 g/kg concentrate on a dry matter (DM) basis were fed once daily for ad libitum intake. The haylage component constituted the dietary treatments: (i) Agriculture and Agri-Food Canada F70 fenugreek (F70), (ii) Crop Development Center Quatro fenugreek (QUAT) and (iii) alfalfa (ALF). DM intake (DMI), milk yield and milk protein and lactose yields were higher (P < 0.001) for cows fed ALF than fenugreek (FEN, average of F70 and QUAT). Milk fat of cows fed FEN contained lower concentrations of saturated, medium-chain and hypercholestrolemic fatty acids (FAs; P < 0.05) than that of cows fed ALF. Apparent total tract digestibility of DM and nutrients was not affected by treatments. Similarly, individual ruminal volatile FA concentrations and rumen pH (5.9) were not affected by treatments. Rumen ammonia-N concentration was higher for FEN than ALF (P < 0.001). Estimates of neutral detergent fiber (NDF) passage rate (P < 0.05) and NDF turnover rate (P < 0.001) in the rumen were higher for ALF than FEN. Our results suggest that although the digestibility of the FEN diets was not different from that of the ALF diet, fenugreek haylage has a lower feeding value than ALF for lactating dairy cows due in part to lower DMI and subsequently lower milk yield.     

The Effects of a Probiotic Yeast on the Bacterial Diversity and Population Structure in the Rumen of Cattle

It has been suggested that the ability of live yeast to improve milk yield and weight gain in cattle is because the yeast stimulates bacterial activity within the rumen. However it remains unclear if this is a general stimulation of all species or a specific stimulation of certain species. Here we characterised the change in the bacterial population within the rumen of cattle fed supplemental live yeast. Three cannulated lactating cows received a daily ration (24 kg/d) of corn silage (61% of DM), concentrates (30% of DM), dehydrated alfalfa (9% of DM) and a minerals and vitamins mix (1% of DM). The effect of yeast (BIOSAF SC 47, Lesaffre Feed Additives, France; 0.5 or 5 g/d) was compared to a control (no additive) in a 3×3 Latin square design. The variation in the rumen bacterial community between treatments was assessed using Serial Analysis of V1 Ribosomal Sequence Tag (SARST-V1) and 454 pyrosequencing based on analysis of the 16S rRNA gene. Compared to the control diet supplementation of probiotic yeast maintained a healthy fermentation in the rumen of lactating cattle (higher VFA concentration [high yeast dose only], higher rumen pH, and lower Eh and lactate). These improvements were accompanied with a shift in the main fibrolytic group (Fibrobacter and Ruminococcus) and lactate utilising bacteria (Megasphaera and Selenomonas). In addition we have shown that the analysis of short V1 region of 16s rRNA gene (50–60 bp) could give as much phylogenetic information as a longer read (454 pyrosequencing of 250 bp). This study also highlights the difficulty of drawing conclusions on composition and diversity of complex microbiota because of the variation caused by the use of different methods (sequencing technology and/or analysis).     

Composition, spatial distribution, and diversity of the bacterial communities in the rumen of cows fed different forages

The species composition, distribution, and biodiversity of the bacterial communities in the rumen of cows fed alfalfa or triticale were investigated using 16S rRNA gene clone library analyses. The rumen bacterial community was fractionated and analyzed as three separate fractions: populations in the planktonic, loosely attached to rumen digesta particles, and tightly attached to rumen digesta particles. Six hundred and thirteen operational taxonomic units (OTUs) belonging to 32 genera, 19 families, and nine phyla of the domain Bacteria were identified from 1014 sequenced clones. Four hundred and fifty one of the 613 OTUs were identified as new species. These bacterial sequences were distributed differently among the three fractions in the rumen digesta of cows fed alfalfa or triticale. Chao 1 estimation revealed that, in both communities, the populations tightly attached to particulates were more diverse than the planktonic and those loosely attached to particulates. S-Libshuff detected significant differences in the composition between any two fractions in the rumen of cows with the same diet and between the communities fed alfalfa and triticale diets. The species richness estimated for the communities fed alfalfa and triticale is 1027 and 662, respectively. The diversity of the rumen bacterial community examined in this study is greater than previous studies have demonstrated and the differences in the community composition between two high-fiber diets have implications for sample selection for downstream metagenomics applications.     

Effect of condensed tannins on bacterial diversity and metabolic activity in the rat gastrointestinal tract

The effect of dietary condensed tannins (proanthocyanidins) on rat fecal bacterial populations was ascertained in order to determine whether the proportion on tannin-resistant bacteria increased and if there was a change in the predominant bacterial populations. After 3 weeks of tannin diets the proportion of tannin-resistant bacteria increased significantly (P < 0.05) from 0.3% +/- 5.5% to 25.3% +/- 8.3% with a 0.7% tannin diet and to 47.2% +/- 5.1% with a 2% tannin diet. The proportion of tannin-resistant bacteria returned to preexposure levels in the absence of dietary tannins. A shift in bacterial populations was confirmed by molecular fingerprinting of fecal bacterial populations by denaturing gradient gel electrophoresis (DGGE). Posttreatment samples were generally still distinguishable from controls after 3.5 weeks. Sequence analysis of DGGE bands and characterization of tannin-resistant isolates indicated that tannins selected for Enterobacteriaceae and Bacteroides species. Dot blot quantification confirmed that these gram-negative bacterial groups predominated in the presence of dietary tannins and that there was a corresponding decrease in the gram-positive Clostridium leptum group and other groups. Metabolic fingerprint patterns revealed that functional activities of culturable fecal bacteria were affected by the presence of tannins. Condensed tannins of Acacia angustissima altered fecal bacterial populations in the rat gastrointestinal tract, resulting in a shift in the predominant bacteria towards tannin-resistant gram-negative Enterobacteriaceae and Bacteroides species.