Effects of red cabbage extract rich in anthocyanins on rumen fermentation,rumen bacterial community,nutrient digestion,and plasma indices in beef bulls

Dietary anthocyanins (ATH) have probiotic and antioxidant functions in humans. They may also have beneficial impacts on rumen microorganisms and subsequently nutrient digestion in cattle. The experiment aimed to study the effects of dietary red cabbage extract (RCE) rich in ATH on rumen fermentation, rumen bacterial community, and nutrient digestibility in beef bulls. Eight Simmental beef bulls and two RCE levels (0 and 120 g/d) were allocated in a replicated 2 × 2 crossover design. Each experimental period included 15 days for adaptation and subsequent 5 days for sampling. The results showed that dietary addition of RCE increased the ruminal concentration of total volatile fatty acids and the molar proportion of propionate, decreased the acetate to propionate ratio, and tended to decrease the molar proportion of acetate, but it did not affect the ruminal pH and the concentrations of ammonia N, microbial CP, monophenols, polyphenols, and total phenolics. ATH was undetectable in the ruminal fluid of beef bulls in both groups. RCE did not affect the alpha diversity of rumen bacterial community, and the relative abundances of major rumen bacteria at the phylum level, but it increased the relative abundances of Ruminobacter and Anaerovibrio and tended to increase the relative abundances of Oribacterium and Monoglobus at the genus level. RCE tended to increase the plasma concentrations of globulin and total protein, but it did not affect the plasma albumin, urea, triglyceride, glucose, and antioxidant activities. Dietary addition of RCE did not affect the apparent nutrient digestibility. In conclusion, the ATH in RCE was highly hydrolysable in rumen fluid. Dietary addition of RCE increased the ruminal concentration of total volatile fatty acids, decreased the acetate to propionate ratio, and slightly modified the rumen bacterial community, but it did not affect the nutrient digestibility and the plasma antioxidants in beef bulls.     

Decreasing the Roughage: Concentrate Ratio of a Diet to Determine the Critical Roughage Part for Beef Cattle

The critical roughage part (CRP) of 2 diet types was determined in a cross-over design with 6 double-muscled and 6 normally conformated Belgian Blue bulls fitted with rumen cannulae. The roughage:concentrate ratio was lowered weekly until signs of a lack of physical structure were observed. For diet 1, consisting of maize silage and concentrates, the initial proportion of maize silage was 25% of DM but it decreased weekly with 5% units of DM. For the second diet, consisting of wheat straw and concentrate, 12% straw (DM basis) was provided during the first week and thereafter the proportion of straw decreased weekly with 3% units of DM. Several directly observable parameters (rumen pH, feed intake, bloat, faecal consistency) were evaluated weekly for each bull. Apart from these direct indicators of acidosis, also other parameters, whose results were only available after the end of the trial, were determined (volatile fatty acid profile, lactic acid concentration, chewing time). The roughage part between the part fed when signs of a lack of physical structure was first observed and the part that was fed the week before, was considered as the CRP. Most animals showed no acute signs of clinical acidosis (directly observable parameters) and finished the trial on a 100% concentrate diet. However, in sacco rumen DM-degradabilities of maize silage, grass silage and wheat grain was depressed considerably when low roughage diets were fed. Based on all observed parameters, the mean CRP was calculated to be 14.7% for diet 1 and 8.1% for diet 2. The beef type (double-muscled or not) had no influence on the CRP.     

Studies on potential effects of fumaric acid on rumen microbial fermentation,methane production and microbial community

The greenhouse gas methane (CH₄) contributes substantially to global climate change. As a potential approach to decrease ruminal methanogenesis, the effects of different dosages of fumaric acid (FA) on ruminal microbial metabolism and on the microbial community (archaea, bacteria) were studied using a rumen simulation technique (RUSITEC). FA acts as alternative hydrogen acceptor diverting 2H from methanogenesis of archaea towards propionate formation of bacteria. Three identical trials were conducted with 12 fermentation vessels over a period of 14 days. In each trial, four fermentation vessels were assigned to one of the three treatment groups differing in FA dosage: low fumaric acid (LFA), high fumaric acid (HFA) and without FA (control). FA was continuously infused with the buffer. Grass silage and concentrate served as substrate. FA led to decreases in pH and to higher production rates of total short chain fatty acids (SCFA) mediated by increases in propionate for LFA of 1.69 mmol d^?1 and in propionate and acetate production for HFA of 4.49 and 1.10 mmol d^?1, respectively. Concentrations of NH₃-N, microbial crude protein synthesis, their efficiency, degradation of crude nutrients and detergent fibre fraction were unchanged. Total gas and CH₄ production were not affected by FA. Effects of FA on structure of microbial community by means of single strand conformation polymorphism (SSCP) analyses could not be detected. Given the observed increase in propionate production and the unaffected CH₄ production it can be supposed that the availability of reduction equivalents like 2H was not limited by the addition of FA in this study. It has to be concluded from the present study that the application of FA is not an appropriate approach to decrease the ruminal CH₄ production.     

Herbage utilisation method affects rumen fluid and milk fatty acid profile in Holstein and Montbéliarde cows

Compared with maize silage- and concentrate-based diets, herbage-based diets were repeatedly shown to favourably influence the milk fatty acid (FA) profile. However, it is unclear how the herbage feeding mode (grazing vs indoor green-feeding) and conservation (fresh herbage vs hay vs silage) modify the milk FA profile. Therefore, the aim of the present experiment was to investigate the effect of different herbage utilisation methods (including herbage feeding mode and herbage conservation method) on the ruminal biohydrogenation of dietary FA and the consequences on the milk FA composition in cows of two breeds (Holstein and Montbéliarde). Concomitant effects of botanical composition and phenological stage of the herbage on milk FA profile were controlled for by harvesting barn-dried hay and silage simultaneously as first cut from the same ryegrass-dominated grassland in a semi-mountainous region. Seven weeks later, the first regrowth of the same plot was used as fresh herbage, either grazed or fed indoor (indoor green-feeding). Twenty-four Montbéliarde and 24 Holstein cows were randomly allocated to four groups of 12 cows balanced by breed, parity, and milk yield. In a free-stall barn, three groups were given ad libitum access to hay, silage, or fresh herbage, respectively. The fourth group was strip-grazing. All cows were supplemented with 3 kg DM/day of the same energy-rich concentrate. After 2 weeks of adaptation to the forage, samples of forage, concentrate, milk, blood, and rumen fluid were collected. Fatty acid composition of forages, rumen fluid, and milk was analysed by gas chromatography. Haymaking reduced total FA content of the herbage, in particular that of linoleic acid (LA) and α-linolenic acid (ALA). Still, rumen fluid lipids of hay-fed cows had the highest proportion of rumenic acid, LA, ALA, and total polyunsaturated fatty acids (PUFAs). Milk fat from hay-fed cows had the highest proportion of LA, and the apparent transfer rates from feed to milk of LA and ALA were higher in hay-fed cows than in silage-fed cows. The proportion of PUFAs was highest in milk fat from grazing and indoor green-fed Montbéliarde cows and lowest in silage-fed cows of both breeds. In conclusion, the herbage utilisation method affects the ruminal biohydrogenation of LA and ALA, whereby herbage drying particularly increases their transfer from herbage to milk.     

Evaluation of the effects of different diets on microbiome diversity and fatty acid composition of rumen liquor in dairy goat

Fat supplementation plays an important role in defining milk fatty acids (FA) composition of ruminant products. The use of sources rich in linoleic and α-linolenic acid favors the accumulation of conjugated linoleic acids isomers, increasing the healthy properties of milk. Ruminal microbiota plays a pivotal role in defining milk FA composition, and its profile is affected by diet composition. The aim of this study was to investigate the responses of rumen FA production and microbial structure to hemp or linseed supplementation in diets of dairy goats. Ruminal microbiota composition was determined by 16S amplicon sequencing, whereas FA composition was obtained by gas-chromatography technique. In all, 18 pluriparous Alpine goats fed the same pre-treatment diet for 40±7 days were, then, arranged to three dietary treatments consisting of control, linseed and hemp seeds supplemented diets. Independently from sampling time and diets, bacterial community of ruminal fluid was dominated by Bacteroidetes (about 61.2%) and Firmicutes (24.2%) with a high abundance of Prevotellaceae (41.0%) and Veillonellaceae (9.4%) and a low presence of Ruminococcaceae (5.0%) and Lachnospiraceae (4.3%). Linseed supplementation affected ruminal bacteria population, with a significant reduction of biodiversity; in particular, relative abundance of Prevotella was reduced (−12.0%), whereas that of Succinivibrio and Fibrobacter was increased (+50.0% and +75.0%, respectively). No statistically significant differences were found among the average relative abundance of archaeal genera between each dietary group. Moreover, the addition of linseed and hemp seed induced significant changes in FA concentration in the rumen, as a consequence of shift from C18 : 2n-6 to C18 : 3n-3 biohydrogenation pathway. Furthermore, dimethylacetal composition was affected by fat supplementation, as consequence of ruminal bacteria population modification. Finally, the association study between the rumen FA profile and the bacterial microbiome revealed that Fibrobacteriaceae is the bacterial family showing the highest and significant correlation with FA involved in the biohydrogenation pathway of C18 : 3n-3.     

Ruminal fermentation characteristics and microbial nitrogen assimilation in sheep fed differently composed grass silages

The investigation aimed at examining if the composition of grassland silage affects the microbial nitrogen assimilation in the rumen of sheep. The silages were made of vegetative summer re-growths consisting of 48% grasses, 28% legumes and 24% other forbs (GCF) or of pure grass (G). Silage GCF contained more intermediately degradable non-structural and less slowly degradable carbohydrates, more crude protein (CP), a narrower ratio between slow and very slow degradable nitrogen (N), and exhibited higher in situ degradability of organic matter and CP than Silage G. Four adult wethers equipped with rumen fistulae were used in a two factorial trial. Feed was offered either as silage alone or as a mixture of silage and barley (60:40). Microbial N was estimated using continuous intraruminal 15N infusion and measurement of 15N-enrichment in microbes isolated from rumen liquor samples. With the exception of trends for ruminal butyrate concentrations, no interactions were detected between silage and barley feeding. Sheep receiving Silage GCF exhibited larger diurnal fluctuations of ammonia, and produced more microbial N (p < 0.05) than sheep on Silage G. Feeding the silages with barley decreased ruminal pH and elevated the concentrations of butyrate (p < 0.05). The 15N incorporation into microbial N was reduced by barley feeding (p < 0.05) along with a trend to accelerated rumen fluid turnover, resulting in similar microbial N yields as found in sheep receiving silage without barley. It is concluded that the larger and better balanced amounts of intermediately degradable carbohydrate- and N-containing fractions favoured the ruminal microbial protein synthesis in sheep consuming Silage GCF instead of Silage G.     

Pantothenic acid supplementation to support rumen microbes?

Based on repeatedly reported extensive pantothenic acid disappearance in the rumen, the present study is aimed at examining if pantothenic acid is used for a more efficient ruminal fermentation and microbial growth in an artificial rumen (Rusitec). Three substrates differing in roughage/concentrate ratio were incubated with and without the addition of Ca-D-pantothenate. Pantothenic acid was extensively degraded without notably influencing fermentation, microbial protein synthesis and the status of other B-vitamins such as riboflavin, vitamin B6 and niacin. Therefore, pantothenic acid supplementation cannot be expected to contribute to microbial benefit for the ruminant animal.     

Nutritional properties of potato protein concentrate compared with soybean meal as the main protein source in feed for the double-muscled Belgian Blue bull

The objective of this experiment was to compare the nutritional properties of potato protein concentrate, a by-product of the starch industry produced entirely in Europe, with that of soybean meal (SBM), for growing cattle. The experiment was conducted on double-muscled Belgian Blue bulls, fitted with rumen, duodenal and ileal cannulas, according to a 4 × 4 Latin square design. They were fed three different iso-N and iso-net energy diets formulated according to the Dutch feed evaluation system, differing in the nature of the main protein source, which was either SBM (‘SBM’ treatment), potato protein concentrate (PPC, ‘PPC’ treatment) or an iso-N mixture of these two protein sources (‘mixed’ treatment). A fourth treatment consisted of ‘PPC’ supplemented by 9.5% digestible proteins supplied by duodenal perfusion of sodium caseinate (CAS, ‘PPC + CAS’ treatment). No significant difference was observed in the ruminal fluid pH, whereas both ‘PPC’ and ‘PPC + CAS’ had the effect of reducing the ruminal ammonia nitrogen (N-NH₃) concentration. No significant difference was observed in the apparent intestinal digestibility of the dry matter (DM), organic matter (OM) or N. Outflows of non-NH₃-N, microbial proteins and dietary proteins from the rumen were similar for ‘PPC’, ‘SBM’ and ‘mixed’, and increased with CAS infusion by 20%, 17% and 27%, respectively. On the basis of in vivo observations, the degradability of SBM and PPC proteins was estimated at 0.60 and 0.43, respectively, corresponding to the values quoted in the literature. The supply of digestible essential amino acids (EAA) was significantly greater with ‘PPC + CAS’ and did not differ among ‘SBM’, ‘mixed’ and ‘PPC’. This illustrates the difficulty of altering the amino acid (AA) pattern of digestible protein by the nature of the protein of dietary origin when an animal is fed a high nutritional value diet. N retention was not affected by replacing SBM with PPC, but increased by 10% with CAS infusion. On the basis of the plasma AA pattern, the supply of digestible Met was probably limiting with ‘SBM’, ‘mixed’ and ‘PPC’. The CAS perfusion supplemented all AA, including Met, leading to increased N retention. This improvement was limited, however, and N utilisation remained unchanged between treatments. In conclusion, despite a more favourable EAA pattern, PPC offered no advantage compared with SBM for growing bulls when diets were formulated according to the Dutch feed evaluation system.     

An in vitro evaluation of browser and grazer fermentation efficiency and microbiota using European moose spring and summer foods

Evolutionary morphological and physiological differences between browsers and grazers contribute to species‐specific digestion efficiency of food resources. Rumen microbial community structure of browsers is supposedly adapted to characteristic nutrient composition of the diet source. If this assumption is correct, domesticated ruminants, or grazers, are poor model animals for assessing the nutritional value of food consumed by browsing game species. In this study, typical spring and summer foods of the European moose (Alces alces) were combined with rumen fluid collected from both dairy cows (Bos taurus) and from moose, with the aim of comparing fermentation efficiency and microbial community composition. The nutritional value of the food resources was characterized by chemical analysis and advanced in vitro measurements. The study also addressed whether or not feed evaluation based on in vitro techniques with cattle rumen fluid as inoculum could be a practical alternative when evaluating the nutritional value of plants consumed by wild browsers. Our results suggest that the fermentation characteristics of moose spring and summer food are partly host‐specific and related to the contribution of the bacterial phyla Firmicutes and Bacteriodetes to the rumen microbial community. Host‐specific adaptations of the ruminal microbial community structure could be explained from the evolutionary adaptations related to feeding habitats and morphophysiological differences between browsers and grazers. However, the observed overall differences in microbial community structure could not be related to ruminal digestion parameters measured in vitro. The in vitro evaluation of digestion efficiency reveals that equal amounts of methane were produced across all feed samples regardless of whether the ruminal fluid was from moose or dairy cow. The results of this study suggested that the nutritional value of browsers' spring and summer food can be predicted using rumen fluid from domesticated grazers as inoculum in in vitro assessments of extent of digestion when excluding samples of the white water lily root, but not of fermentation characteristics as indicated by the proportions of individual fermentation fatty acids to the total of volatile fatty acids.     

Oral Samples as Non-Invasive Proxies for Assessing the Composition of the Rumen Microbial Community

Microbial community analysis was carried out on ruminal digesta obtained directly via rumen fistula and buccal fluid, regurgitated digesta (bolus) and faeces of dairy cattle to assess if non-invasive samples could be used as proxies for ruminal digesta. Samples were collected from five cows receiving grass silage based diets containing no additional lipid or four different lipid supplements in a 5 x 5 Latin square design. Extracted DNA was analysed by qPCR and by sequencing 16S and 18S rRNA genes or the fungal ITS1 amplicons. Faeces contained few protozoa, and bacterial, fungal and archaeal communities were substantially different to ruminal digesta. Buccal and bolus samples gave much more similar profiles to ruminal digesta, although fewer archaea were detected in buccal and bolus samples. Bolus samples overall were most similar to ruminal samples. The differences between both buccal and bolus samples and ruminal digesta were consistent across all treatments. It can be concluded that either proxy sample type could be used as a predictor of the rumen microbial community, thereby enabling more convenient large-scale animal sampling for phenotyping and possible use in future animal breeding programs aimed at selecting cattle with a lower environmental footprint.     

Influence of various amounts of fumaric acid on performance and parameters of the acid-base balance of growing bulls fed with grass or maize silage

The aim of the present study was to determine the effects of the potential methane reducer fumaric acid on the fattening performance and acid-base balance of growing bulls fed two different silage types as roughage (maize and grass silage). A total of 62 fattening bulls (German Holstein breed, initial body weight: 266 +/- 42 kg), randomly assigned to eight feeding groups, received four levels of fumaric acid (0, 100, 200 or 300 g/d) at each silage type. The daily feed and water intake and the live weight were measured over the whole testing period of 280 days. In blood samples, blood cells and blood gases as a parameter of acid-base status were analysed. Feed and faeces were collected to determine the apparent nutrient digestibility. Five animals from each group fed maize or grass silage, supplemented with 0 g or 300 g fumaric acid, were slaughtered at 580 kg body weight. After slaughter, rumen fluid pH was measured and dressing percentage was calculated. Neither the total feed intake (8.81 +/- 0.07 kg/d) nor the daily weight gain (1277 +/- 24 g/d) was influenced significantly by treatments. Fumaric acid supplementation did not influence the erythrocyte count or the blood gas concentration. The silage type significantly influenced the apparent digestibility of the whole diet. The dressing percentage was slightly higher (p < 0.1) after fumaric acid supplementation. No signs of an incompatibility to fumaric acid on the animals were observed over the whole experimental period. However, it seems to be necessary to conduct more long-term studies with different silage types and addition of organic acids combined with direct measurements of methane.     

Evaluation of the effects of dietary particle fractions on fermentation profile and concentration of microbiota in the rumen of dairy cows fed grass silage-based diets

The study evaluated the effects of three different theoretical particle lengths (TPL) of grass silage on the distribution of particle fractions of the diet and the resulting effects on fermentation profile and concentrations of protozoa and mixed bacterial mass in the rumen of three lactating Holstein cows fed total mixed rations (45% grass silage, 5% grass hay and 50% concentrate) ad libitum. Decreasing TPL of grass silage (long, medium, short) reduced particles retained on the 19-mm sieve of the Penn State Particle Separator, while particle fractions from 8 mm to 19 mm and smaller than 8 mm were increased. Different TPL did not affect pH and the concentration of volatile fatty acids in the rumen. However, lowering the TPL from long to medium increased significantly the bicarbonate concentration, acetate proportion and protozoal number in the rumen, whereas the proportion of bacterial protein in ruminal digesta and its amino acid concentration were significantly increased by the short TPL. For the current feeding conditions, it can be concluded that increasing the fraction of particles between 8 and 19 mm and probably even the fraction below 8 mm by decreasing TPL of grass silage do not adversely affect rumen conditions and can be beneficial in terms of optimising concentration and activity of ruminal microbiota in high-yielding dairy cows.     

Rumen fermentation,nutrient flow to the duodenum and digestibility in bulls fed calcium soaps of rapeseed fatty acids and soya bean meal coated with calcium soaps

Three Polish Friesian bulls fitted with rumen and duodenal canulas were used in a 3 × 3 Latin square experiment to study the effect of Ca soaps of rapeseed fatty acids (CSRFA) on rumen fermentation, nutrient and fatty acid (FA) flow to the duodenum, and to investigate in vivo the possibility of using CSRFA as a means of protecting soya bean meal protein against degradation in the rumen. Treatments were (1) control, no fat (2) CSRFA, at 2% of dietary DM and (3) SBM protein coated with CSRFA (CSRFA:SBM, 1:1, wt/wt), at 4% of dietary DM. The animals were fed isonitrogenous diets, containing 45% meadow hay, 10% fodder beet and 45% concentrate mixture on a DM basis. Intake was limited to 80 g DM d⁻¹ kg^−0.75. There were no treatment effects on rumen fermentation, rumen liquid turnover rate or volume, suggesting that CSRFA were inert in the rumen. A slight decrease in ammonia concentration in the rumen fluid from the CSRFA:SBM diet and the degradability data do not confirm the protection of SBM protein by CSRFA. Treatments did not affect apparent OM digestibility in the rumen, total N and NAN (non-ammonia nitrogen) flow into the duodenum or microbial protein synthesis. However, rumen degradability of protein was increased by feeding CSRFA. Feeding CSRFA significantly increased the duodenal flows of C18:0, C18:1 and total FA, but when expressed as a percentage of intake, there were no significant treatment differences. The average duodenal flow of total FA was 81.7% of intake. Extents of biohydrogenation of unsaturated FA were decreased in diets containing CSRFA. Estimated net biohydrogenation of C18:2, C18:3 and total unsaturated C18 FA of CSRFA was 43.3, 60.4 and 59.4%, respectively. Postrumen and total tract digestibility of total and individual FA were not different between treatments     

Evaluation of the effects of dietary particle fractions on fermentation profile and concentration of microbiota in the rumen of dairy cows fed grass silage-based diets

The study evaluated the effects of three different theoretical particle lengths (TPL) of grass silage on the distribution of particle fractions of the diet and the resulting effects on fermentation profile and concentrations of protozoa and mixed bacterial mass in the rumen of three lactating Holstein cows fed total mixed rations (45% grass silage, 5% grass hay and 50% concentrate) ad libitum. Decreasing TPL of grass silage (long, medium, short) reduced particles retained on the 19-mm sieve of the Penn State Particle Separator, while particle fractions from 8 mm to 19 mm and smaller than 8 mm were increased. Different TPL did not affect pH and the concentration of volatile fatty acids in the rumen. However, lowering the TPL from long to medium increased significantly the bicarbonate concentration, acetate proportion and protozoal number in the rumen, whereas the proportion of bacterial protein in ruminal digesta and its amino acid concentration were significantly increased by the short TPL. For the current feeding conditions, it can be concluded that increasing the fraction of particles between 8 and 19 mm and probably even the fraction below 8 mm by decreasing TPL of grass silage do not adversely affect rumen conditions and can be beneficial in terms of optimising concentration and activity of ruminal microbiota in high-yielding dairy cows.     

Effect of chitosans on in vitro rumen digestion and fermentation of maize silage

The gas in vitro technique was used to study the effects of six types of chitosans, each having different molecular weights and acetylation degrees, on rumen microbial fermentation. In a first trial, a separate concentration of 750 mg/l of culture fluid for each of the six chitosans (CHI1, CHI2, CHI3, CHI4, CHI5, and CHI6) was incubated for 24 h in diluted ruminal fluid with maize silage as the substrate. The ionophore antibiotic monensin (MON) was used as a positive control, and a negative control with no chitosan (CTR) was also included. Each treatment was tested in triplicate for three different periods. At the end of the trial, samples were collected to determine volatile fatty acid (VFA) and ammonia N concentrations, and pH and gas production values were recorded. Methane concentration was estimated stoichiometrically. In vitro true organic matter digestibility (IVOMD) and partitioning factor (PF, mg OM truly degraded/ml gas produced) were also calculated. In a second trial, a separate concentration of 750 mg/l of each of the six chitosans was incubated for 144 h in diluted ruminal fluid with maize silage as the substrate, to study the effects of these compounds on fermentation kinetics.All six chitosans decreased the IVOMD and PF values. Chitosan inclusion did not affect the fermentation of the substrate's soluble fraction, but did reduce the fermentation kinetics of the insoluble but fermentable fraction. However, only CHI5 and CHI6 decreased total VFA concentration. CHI3 and CHI6 decreased the molar proportion of acetate and increased the molar proportion of propionate, thus increasing the propionate-to-acetate ratio. Chitosan inclusion did not affect molar proportions of butyrate. With the exception of CHI2, the molar proportion of branch-chained VFA was lowered by all of the chitosan treatments. Most of the treatments also decreased methane production, also with the exception of CHI2.In conclusion, chitosan extracts may enable the manipulation of rumen microbial fermentation, but further research is required to elucidate the effect of chitosans on ruminal fermentation parameters in commercial diets as well as the adaptability of rumen microflora to these additives.     

Influence of various amounts of fumaric acid on performance and parameters of the acid–base balance of growing bulls fed with grass or maize silage

The aim of the present study was to determine the effects of the potential methane reducer fumaric acid on the fattening performance and acid–base balance of growing bulls fed two different silage types as roughage (maize and grass silage). A total of 62 fattening bulls (German Holstein breed, initial body weight: 266 ± 42 kg), randomly assigned to eight feeding groups, received four levels of fumaric acid (0, 100, 200 or 300 g/d) at each silage type. The daily feed and water intake and the live weight were measured over the whole testing period of 280 days. In blood samples, blood cells and blood gases as a parameter of acid–base status were analysed. Feed and faeces were collected to determine the apparent nutrient digestibility. Five animals from each group fed maize or grass silage, supplemented with 0 g or 300 g fumaric acid, were slaughtered at 580 kg body weight. After slaughter, rumen fluid pH was measured and dressing percentage was calculated. Neither the total feed intake (8.81 ± 0.07 kg/d) nor the daily weight gain (1277±24 g/d) was influenced significantly by treatments. Fumaric acid supplementation did not influence the erythrocyte count or the blood gas concentration. The silage type significantly influenced the apparent digestibility of the whole diet. The dressing percentage was slightly higher (p < 0.1) after fumaric acid supplementation. No signs of an incompatibility to fumaric acid on the animals were observed over the whole experimental period. However, it seems to be necessary to conduct more long-term studies with different silage types and addition of organic acids combined with direct measurements of methane.     

Branched-chain volatile fatty acids and folic acid accelerated the growth of Holstein dairy calves by stimulating nutrient digestion and rumen metabolism

The combined addition of branched-chain volatile fatty acids (BCVFAs) and folic acid (FA) could improve growth performance and nutrient utilization by stimulating ruminal microbial growth and enzyme activity. This study was conducted to evaluate the effects of BCVFA and FA addition on growth performance, ruminal fermentation, nutrient digestibility, microbial enzyme activity, microflora and excretion of urinary purine derivatives (PDs) in calves. Thirty-six Chinese Holstein weaned calves (60 ± 5.4 days of age and 107 ± 4.7 kg of BW) were assigned to one of four groups in a randomized block design. Treatments were control (without additives), FA (with 10 mg FA/kg dietary DM), BCVFA (with 5 g BCVFA/kg dietary DM) and the combined addition of FA and BCVFA (10 mg/kg DM of FA and 5 g/kg DM of BCVFA). Supplements were hand-mixed into the top one-third of total mixed ration. Dietary concentrate to maize silage ratio was 50 : 50 on a DM basis. Dietary BCVFA or FA addition did not affect dry matter intake but increased average daily gain (ADG) and feed conversion efficiency. Ruminal pH and ammonia N were lower, and total volatile fatty acids (VFAs) concentration was higher for BCVFA or FA addition than for control. Dietary BCVFA or FA addition did not affect acetate proportion but decreased propionate proportion and increased acetate to propionate ratio. Total tract digestibility of DM, organic matter, CP and NDF was higher for BCVFA or FA addition than for control. Dietary BCVFA or FA addition increased activity of carboxymethyl cellulase and cellobiase, population of total bacteria, fungi, Ruminococcus albus, R. flavefaciens, Fibrobacter succinogenes and Prevotella ruminicola as well as total PD excretion. Ruminal xylanase, pectinase and protease activity and Butyrivibrio fibrisolvens population were increased by BCVFA addition, whereas population of protozoa and methanogens was increased by FA addition. The BCVFA × FA interaction was significant for acetate to propionate ratio, cellobiase activity and total PD excretion, and these variables increased more with FA addition in diet without BCVFA than in diet with BCVFA. The data indicated that supplementation with BCVFA or FA increased ADG, nutrient digestibility, ruminal total VFA concentration and microbial protein synthesis by stimulating ruminal microbial growth and enzyme activity in calves.     

Metagenomic analysis of Surti buffalo (Bubalus bubalis) rumen: a preliminary study

The complex microbiome of the rumen functions as an effective system for the conversion of plant cell wall biomass to microbial proteins, short chain fatty acids and gases. In this study, metagenomic approaches were used to study the microbial populations and metabolic potential of the microbial community. DNA was extracted from Surti Buffalo rumen samples (four treatments diet) and sequenced separately using a 454 GS FLX Titanium system. We used comparative metagenomics to examine metabolic potential and phylogenetic composition from pyrosequence data generated in four samples, considering phylogenetic composition and metabolic potentials in the rumen may remarkably be different with respect to nutrient utilization. Assignment of metagenomic sequences to SEED categories of the Metagenome Rapid Annotation using Subsystem Technology (MG-RAST) server revealed a genetic profile characteristic of fermentation of carbohydrates in a high roughage diet. The distribution of phylotypes and environmental gene tags (EGTs) detected within each rumen sample were dominated by Bacteroidetes/Chlorobi, Firmicutes and Proteobacteria in all the samples. The results of this study could help to determine the role of rumen microbes and their enzymes in plant polysaccharide breakdown is fundamental to understanding digestion and maximising productivity in ruminant animals.     

Effect of different levels of concentrate on ruminal microorganisms and rumen fermentation in Nellore steers

The aim of this study was to investigate the effect of different dietary levels of concentrate on feed intake, digestibility, ruminal fermentation and microbial population in steers. Eight Nellore steers fitted with ruminal cannulas were used in a double 4 × 4 Latin square design experiment. The dietary treatments consist of four different proportions of concentrate to roughage: 30:70, 40:60, 60:40 and 80:20% in the dry matter, resulting in Diets 30, 40, 60 and 80, respectively. The roughage was corn silage, and the concentrate was composed of corn, soybean meal and urea. Apparent digestibility of organic matter and crude protein showed a linear association with concentrate proportion (p = 0.01), but the increased concentrate levels did not affect the digestibility of fibre. The lowest ruminal pH-values were observed in animals fed with Diet 80, remaining below pH 6.0 from 6 h after feeding, while in the other diets, the ruminal pH was below 6.0 not before 12 h after feeding. After feeding Diet 80, the ammonia concentration in the rumen was significantly the highest. Higher dietary concentrate levels resulted in a linear increase of propionic acid concentrations, a linear reduction of the ratio acetic acid to propionic acid (p < 0.01) and a linear increased synthesis of microbial nitrogen (p < 0.001). The predicted production of methane was lower in diets with greater amounts of concentrate (p = 0.032). The population of methanogens, R. flavefaciens and R. albus decreased with higher concentrate levels, while the population of S. ruminantium increased (p < 0.05). The results indicate that greater amounts of concentrate do not decrease ruminal pH-values as much as expected and inhibit some cellulolytic bacteria without impairing the dry matter intake and fibre digestibility in Nellore steers.     

Evaluation of group-specific, 16S rRNA-targeted scissor probes for quantitative detection of predominant bacterial populations in dairy cattle rumen

AIMS: To develop a suite of group-specific, rRNA-targeted oligonucleotide scissor probes for the quantitative detection of the predominant bacterial groups within the ruminal microbial community with the rRNA cleavage reaction-mediated microbial quantification method. METHODS AND RESULTS: Oligonucleotides that complement the conserved sites of the 16S rRNA of phylogenetically defined groups of bacteria that significantly contribute to the anaerobic fermentation of carbohydrates in ruminal ecosystems were selected from among published probes or were newly designed. For each probe, target-specific rRNA cleavage was achieved by optimizing the formamide concentration in the reaction mixture. The set of scissor probes was then used to analyse the bacterial community in the rumen fluids of four healthy dairy cows. In the rumen fluid samples, the genera Bacteroides/Prevotella and Fibrobacter and the Clostridium coccoides-Eubacterium rectale group were detected in abundance, accounting for 44-48%, 2.9-10%, and 9.1-10% of the total 16S rRNA, respectively. The coverage with the probe set was 71-78% of the total bacterial 16S rRNA. CONCLUSIONS: The probe set coupled with the sequence-specific small-subunit rRNA cleavage method can be used to analyse the structure of a ruminal bacterial community. SIGNIFICANCE AND IMPACT OF THE STUDY: The probe set developed in this study provides a tool for comprehensive rRNA-based monitoring of the community members that dominate ruminal ecosystems. As the ruminal microbial community can be perturbed, it is important to track its dynamics by analysing microbiological profiles under specific conditions. The method described here will provide a convenient approach for such tracking.