Partial characterization of phylogeny,ecology and function of the fibrolytic bacterium Ruminococcus flavefaciens OS14, newly isolated from the rumen of swamp buffalo

The fibrolytic rumen bacterium Ruminococcus flavefaciensOS14 was isolated from swamp buffalo and its phylogenetic, ecological and digestive properties were partially characterized. Isolates from rumen contents of four swamp buffalo were screened for fibrolytic bacteria; one of the 40 isolates showed a distinctive feature of solubilizing cellulose powder in liquid culture and was identified as R. flavefaciens based on its 16S ribosomal DNA sequence. This isolate, OS14, was employed for detection and digestion studies, for which a quantitative PCR assay was developed and defined cultures were tested with representative forages in Thailand. OS14 was phylogenetically distant from other isolated and uncultured R. flavefaciens and showed limited distribution among Thai ruminants but was absent in Japanese cattle. OS14 digested rice straw and other tropical forage to a greater extent than the type strain C94 of R. flavefaciens. OS14 produced more lactate than C94, and digested para grass to produce propionate more extensively in co‐culture with lactate‐utilizing Selenomonas ruminantium S137 than a co‐culture of C94 with S137. These results indicate that phylogenetically distinct OS14 could digest Thai local forage more efficiently than the type strain, possibly forming a symbiotic cross‐feeding relationship with lactate‐utilizing bacteria. This strain might be useful for future animal and other industrial applications.     

Number of nitrate- and nitrite-reducing Selenomonas ruminantium in the rumen, and possible factors affecting its growth

The cell number of Selenomonas ruminantium (S. ruminantium) that reduces nitrate and nitrite in the rumen was usually 8–10% of the total number of S. ruminantium (an order of 10⁶/mL). The percentage was not affected by the roughage/concentrate ratio or nitrate content of the diet in 2 weeks. However, feeding a high‐nitrate diet for 12 weeks increased the percentage. The percentage of lactate‐using S. ruminantium, such as the ssp. lactilytica, was less than 1% of the total number of S. ruminantium. No S. ruminantium was found that used formate as an electron donor for nitrate and nitrite reduction. Lactate and H₂ appeared to be important for nitrate and nitrite reduction by S. ruminantium. Nitrate reduction by S. ruminantium was enhanced by the coexistence of amylolytic bacteria in a medium containing starch, and as a result, nitrite accumulation increased. Coexistence of cellulolytic bacteria facilitated the growth of S. ruminantium in a medium containing cellulose, and consequently increased nitrite reduction. In order to suppress nitrite accumulation in the rumen, it may be important to enhance fiber digestion.     

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.     

Interaction of rumen bacteria as assumed by colonization patterns on untreated and alkali‐treated rice straw

Colonization patterns of representative rumen bacteria were compared between untreated rice straw (UTS) and sodium hydroxide‐treated rice straw (SHTS). UTS and SHTS were incubated in the rumen of sheep for 10 min, 1, 2, 6, 12, 24, 48 and 96 h using the nylon bag method. The population sizes of 13 representative bacterial species or groups were quantified by real‐time PCR. The total bacterial population size (abundance) was similar in both UTS and SHTS. Fibrobacter succinogenes showed a higher population size compared to other fibrolytic species and was detected at a higher level in SHTS (3.7%) than in UTS (2.6%). Ruminococcus albus and Ruminococcus flavefaciens were also detected at higher levels in SHTS (0.15% and 0.29%) than in UTS (0.03% and 0.18%). Population sizes of non‐fibrolytic species, such as Selenomonas ruminantium, Anaerovibrio lipolytica and Succinivibrio dextrinosolvens were higher in UTS than in SHTS. Coefficient of determination (r²) on population changes between bacterial species or groups were higher in UTS than in SHTS, suggesting the necessity of stronger bacterial interactions for UTS digestion. Therefore, not only colonization of fibrolytic species, but also synergistic interactions between different bacterial species may be key to the ruminal digestion of rice straw.     

Effect of non‐starch‐polysaccharide‐degrading enzymes as feed additive on the rumen bacterial population in non‐lactating cows quantified by real‐time PCR

The effects of non‐starch‐polysaccharide‐degrading enzymes, added to a maize silage‐ and grass silage‐based total mixed ration (TMR) at least 14 h before feeding, on the rumen bacterial population were investigated. Six non‐lactating Holstein Friesian cows were allocated to three treatment groups using a duplicate 3 × 3 Latin square design with three 31‐day periods (29 days of adaptation and 2 days of sampling). Treatments were control TMR [69% forage and 31% concentrates on a dry matter (DM) basis] or TMR with 13.8 or 27.7 ml/kg of feed DM of Roxazyme G2 liquid with activities (U/ml enzyme preparation) of xylanase 260 000, β‐glucanase 180 000 and cellulase 8000 (DSM Nutritional Products, Basel, Switzerland). The concentrations of 16S rDNA of Anaerovibrio lipolytica, Fibrobacter succinogenes, Prevotella ruminicola, Ruminococcus flavefaciens, Selenomonas ruminantium and Treponema bryantii, and their relative percentage of total bacteria in rumen samples obtained before feeding and 3 and 7 h after feeding and from two rumen fractions were determined using real‐time PCR. Sampling time had only little influence, but bacterial numbers and the composition of the population differed between the transition layer between rumen fluid and the fibre mat (fraction A) and the rumen fluid (fraction B) highlighting the importance to standardize sampling. The 16S rDNA copies of total bacteria and the six bacterial species as well as the population composition were mainly unaffected by the high levels of exogenous enzymes supplemented at all sampling times and in both rumen fractions. Occasionally, the percentages of the non‐fibrolytic species P. ruminicola and A. lipolytica changed in response to enzyme supplementation. Some increases in the potential degradability of the diet and decreases in lag time which occurred collaterally indicate that other factors than changes in numbers of non‐particle‐associated bacteria are mainly responsible for the effects of exogenous enzymes.     

ORIGINAL ARTICLE: Description of development of rumen ecosystem by PCR assay in milk‐fed,weaned and finished lambs in an intensive fattening system

This study examined the reticulo‐rumen characteristics of the microbial community and its fermentative characteristics in milk‐fed, at weaning and finished lambs in a conventional fattening system. Five lambs were assigned to each of three groups: milk‐fed lambs slaughtered at 30 days (T30), weaned lambs slaughtered at 45 days (T45) and ‘finished lambs’ slaughtered at 90 days (T90). At slaughter, rumen size, fermentation parameters (pH, volatile fatty acids and microbial enzyme activity) and protozoal counts were recorded. Quantitative PCR was used to quantify the genes encoding 16S and 18S ribosomal DNA of the rumen bacterial and protozoal populations, respectively, and the sequential colonization of the rumen by cellulolytic (Ruminococcus albus, Ruminococcus flavefaciens) and amylolytic (Prevotella ruminicola, Streptococcus bovis) bacteria, and protozoa (Entodinium sp.). Denaturing gradient gel electrophoresis was used to study the development of rumen microbiota biodiversity. Intake of solid food before weaning caused a significant increase in rumen weight (p < 0.0001) and bacterial DNA (p < 0.05) and volatile fatty acid analysis concentration (p < 0.01), whereas pH declined. In milk‐fed lambs, cellulolytic bacteria were evident after 30 days. Thereafter, in the 45‐day and 90‐day groups, the proportions of R. flavefaciens decreased and R. albus increased. Amylolytic bacteria were present in milk‐fed lambs; the proportion of P. ruminicola increased in fattening lambs and S. bovis was the least abundant species. Protozoal concentrations were irregular; milk‐fed lambs had a significant number of protozoa species from Entodinium and subfamily Isotrichiidae, but they disappeared at weaning. Lamb rumen were refaunated in some individuals at 90 days (Entodinium and subfamily Diplodiniinae spp.), although individual concentrations were variable.     

Effect of α-cyclodextrin-allyl isothiocyanate on ruminal microbial methane production in vitro

The objective of the present study was to investigate the effects of α‐cyclodextrin‐allyl isothiocyanate (CD‐AI) on ruminal microbial methane production and rumen fermentation of corn starch, soluble potato starch or hay plus concentrate (1.5:1) by mixed rumen microorganisms. Diluted rumen fluid (30 mL) was incubated anaerobically at 38°C for 6 and 24 h with or without CD‐AI (0, 0.4, 0.8, 1.6 and 3.2 g/L). The pH of the medium was unchanged by CD‐AI in all substrates. The molar proportion of acetate was decreased and propionate was increased with a corresponding decrease in acetate : propionate ratio (P < 0.05). Total volatile fatty acids and butyrate were increased (P < 0.05). Ammonia‐N was decreased (P < 0.05). Except with soluble potato starch, numbers of protozoa were unchanged after 6 h. As concentration of CD‐AI increased from 0 to 3.2 g/L, fermentation of corn starch, soluble potato starch and hay plus concentrate resulted in decreased (P < 0.05) methane production of 49–100% (6 h) and 14–100% (24 h); 39–100% (6 h) and 16–100% (24 h); and 45–100% (6 h) and 17–100% (24 h), respectively. When hay plus concentrate was used as substrate, methanogenic bacteria were decreased (P < 0.05) with 0.8 g/L of CD‐AI after 6 h. Excluding the lower dose level (0.4 g/L) of CD‐AI, digestibility of neutral detergent fiber of hay plus concentrate was decreased (P < 0.05) after 24 h. A suitable level of CD‐AI could therefore be used as a supplement to inhibit methane production and improve rumen fermentation without detrimental effects on fiber digestion.     

Chemical compositions, feed intakes and digestibilities of crop residue based rations in non-lactating Red Sokoto goats in the subhumid zone of Nigeria

The present study was conducted to evaluate the chemical composition, intake and digestibility of crop residue based rations by Red Sokoto goats maintained on natural pastures and Digitaria smutsii hay during the dry season of the year. Twenty‐eight non‐lactating does were blocked for weight and assigned to 7 treatment groups comprising 4 does each in a completely randomized design. Ration A, the conventional concentrate ration, was used as the positive control, Rations B and C were the two crop residue based test rations, while Ration D, the unsupplemented treatment, was used as the negative control. Each of the supplementation rations was fed at 1% and 2% of the doe's body weight. Ration A had the highest crude protein percentage of 17.19% while Rations B and C had 9.54 and 10.38%, respectively. The naturally grazed pastures and Digitaria smutsii hay (Ration D) contained the least protein: 2.76 and 4.75%, respectively. Ration D also had the highest percentages of acid detergent fiber, neutral detergent fiber and lignin (49.14, 74.73 and 9.49% in hay and 50.29, 8.27 and 11.5% in grazed pastures, respectively). Ration A on the other hand, had the lowest percentages of acid detergent fiber (20.00%), neutral detergent fiber (40.01%) and lignin (4.64%). The results indicated that the supplemented group of does had significantly higher (P < 0.05) dry matter and crude protein intakes as well as nutrient digestibilities than the unsupplemented groups. A comparison of the unsupplemented animals with all the other treatment groups revealed that dry matter digestibility improved by a range of 4.1–27.9%, while crude protein digestibility improved by 17.1–42.2%, the highest value being in does on Ration A. It was concluded that goats were able to subsist and make appreciable gains in the long dry season on crop‐based diets that compared favorably with the conventional concentrate rations. Of the two tested crop residue based rations, Ration C is a better supplementation package than Ration B.     

Effects of cyclodextrin-iodopropane complex on methane production, ruminal fermentation and microbes, digestibility and blood metabolites in steers

The effects when adding cyclodextrin‐iodopropane complex (CD‐IP) to a diet, on ruminal fermentation and microbes, digestibility, blood metabolites and methane production, were evaluated using four Holstein steers in a cross‐over design. The steers were fed Sudangrass hay plus concentrate mixture at a ratio 1.5:1, and CD‐IP (1% of dry matter) was given twice daily by mixing with concentrate mixture. Rumen and blood samples were collected at 0, 2, and 5 h after morning dosing. Ruminal pH and numbers of protozoa were unaffected by CD‐IP treatment. Ruminal molar proportion of acetate was decreased (P < 0.05), and propionate was increased (P < 0.01) at 2 h after CD‐IP dosing. Proportion of butyrate was increased (P < 0.05) and ammonia‐N was decreased (P < 0.05) at 2 and 5 h after CD‐IP dosing. Adding CD‐IP had no effect on the feed intake and digestion of nutrients. Plasma glucose was increased and urea‐N was decreased (P < 0.05) at 2 and 5 h after CD‐IP dosing. Methane production was decreased (P < 0.05) by approximately 18% in the treatment steers. Numbers of methanogenic bacteria were decreased (P < 0.05), while total viable counts, cellulolytic, sulfate reducing and acetogenic bacteria were unaffected. The present results are the first to show that CD‐IP can partially inhibit in vivo ruminal methanogenesis without adverse effects on digestion of nutrients.     

Associative effects of supplementing rice straw‐based diet with cornstarch on intake,digestion, rumen microbes and growth performance of Huzhou lambs

Thirty‐six male Hu lambs consuming a rice straw‐based diet were used in a 60‐day trial to study the associative effects of cornstarch supplementation on intake, digestion, ruminal microbial population and growth performance. All animals were fed rice straw ad libitum together with 160 g/day of rapeseed meal and supplemented with cornstarch at levels of 0 (control), 60, 120 or 180 g/day, respectively. Increment of supplementary cornstarch showed little influence on rice straw intake. Optimal growth performance and highest apparent digestibility of organic matter was achieved in the 120 g/day cornstarch group (P < 0.05), while the digestibilities of neutral detergent fiber and crude protein were significantly decreased by 180 g/day cornstarch (P < 0.05). Similar results were observed for carboxymethyl cellulose activity and relative populations of cellulolytic bacteria (Ruminococcus albus, Ruminococcus flavefaciens and Fibrobacter succinogene). Blood urea nitrogen was reduced by supplementary cornstarch, indicating enhanced protein utilization efficiency. Carcass traits were all significantly improved by supplementary cornstarch. These results suggested that proper amounts of starch supplementation (within 0.5% BW) has little adverse effect on forage utilization, but could effectively improve growth performance. High levels of cornstarch, however, would decrease cellulase activity and populations of cellulolytic bacteria, and hence the digestibility of forage.     

Degradation of cellulose and forage fiber fractions by ruminal cellulolytic bacteria alone and in coculture with phenolic monomer-degrading bacteria.

We hypothesized that bacterial species capable of metabolizing phenolic monomers may act as catalysts for forage fiber breakdown by increasing microbial access to cell wall polysaccharides. Ruminal cellulolytic bacteria alone and in combination with phenolic-degrading bacteria were examined for differences in their ability to degrade fiber fractions of alfalfa or bromegrass. Electron micrographs of Fibrobacter succinogenes S85 cultured in combination with the ruminal phenolic-degrading organisms Eubacterium oxidoreducens G41 and Syntrophococcus sucromutans S195 indicated that bromegrass was degraded more extensively by the triculture than by the monoculture. The sequential detergent system was used to quantify the digestibility of fiber components from alfalfa and bromegrass. F. succinogenes incubated with the two phenolic-degrading organisms did not degrade more cell wall material than did F. succinogenes alone. However, with two other ruminal cellulolytic organisms, Clostridium longisporum B6405 and Ruminococcus albus B6403, greater (P less than .05, P less than .10, respectively) amounts of hemicellulose were degraded (72 h in vitro fermentation) from whole-plant alfalfa when E. oxidoreducens and S. sucromutants were combined with the cellulolytic species than when their monocultures were tested. Similar increases were not observed using a NDF preparation of alfalfa as the substrate. Based on these in vitro experiments, it does not seem that E. oxidoreducens and S. sucromutans play an important role in improving forage fiber degradation by cellulolytic ruminal bacteria.     

Ecological characterization of three different phylogenetic groups belonging to the cellulolytic bacterial species Fibrobacter succinogenes in the rumen

To study the group‐dependent ecology of Fibrobacter succinogenes in the rumen, real‐time polymerase chain reaction assays for two phylogenetic groups (groups 2 and 3) of F. succinogenes were newly established and applied to rumen samples. Both the assays targeting the bacterial 16S rDNA were sensitive and accurate, showing wide quantifiable ranges (10⁴?10⁹ and 10²?10⁹ copies of 16S rDNA) and high recoveries of known amounts of added DNA (96.9 and 98.0%). The quantity of group 1 was confirmed to be numerable by subtracting assay values of groups 2 and 3 from that of F. succinogenes species (groups 1–3). By using the developed assays and the above subtractive calculation, the quantities of all three groups were evaluated in solid and liquid fractions of the rumen content and also on hay stems. In the solid fraction, groups 1 and 2 were abundantly present, compared with group 3 (P < 0.05). On untreated hay stems, group 1 was dominant throughout 48 h. In addition, group 1 showed growth even on the cellulase‐treated hay stems, unlike the other two groups. These results suggest that F. succinogenes group 1 greatly contributes to rumen fiber digestion, even for less degradable materials.     

Diversity of chemical composition, dry matter intake, in vivo digestibility and in situ dry matter degradability of oat hay (Avena sativa)

Diversity of chemical composition, in vivo digestibility and in situ dry matter degradability of oat hay (Avena sativa) were studied. Oat hay samples obtained from 22 batches imported were analyzed for chemical composition. The result showed a wide range of variations in crude protein (3.9–8.8%, on a dry matter basis), crude fiber (22.1–35.2%), nitrogen free extract (50.0–66.0%), organic cellular contents (23.4–40.0%), organic cell wall (53.1–70.8%), organic a (4.1–9.8%), organic b (49.0–63.8%) fractions and the estimated total digestible nutrients (50.3–63.8%) from the organic components. Of these, four oat hay samples were used for the measurement of dry matter intake, in vivo digestibility and in situ dry matter degradability in sheep. Significant differences were observed for the dry matter intake, in vivo digestibility of dry matter, crude protein, crude fiber and nitrogen free extract, dry matter disappearance at 48 h and effective degradability at k = 0.05 h^?1 of the samples (P < 0.05). The results revealed a wide range of diversity of nutritional quality of the imported oat hay that could have an affect on production potential in animals.     

Effect of cyclodextrin diallyl maleate on methane production, ruminal fermentation and microbes in vitro and in vivo

Effects of β‐cyclodextrin diallyl maleate (CD‐M) on methane production, ruminal fermentation and digestibility were studied both in vitro and in vivo. In in vitro study, diluted ruminal fluid (30 mL) was incubated anaerobically at 38°C for 6 and 24 h with or without CD‐M using hay plus concentrate (1.5:1) as a substrate. The CD‐M was added at different concentrations (0, 1.25, 2.5, 5.0 and 7.5 g/L). The pH of the medium and numbers of protozoa were not affected by the addition of CD‐M. Total volatile fatty acids were increased and ammonia‐N was decreased, molar proportion of acetate was decreased and propionate was increased (P < 0.05) by CD‐M. Methane was inhibited (P < 0.05) by 14–76%. The effect of CD‐M on methane production and ruminal fermentation was further investigated in vivo using four Holstein steers in a cross‐over design. The steers were fed Sudangrass hay and concentrate mixture (1.5:1) with or without CD‐M (2% of feed dry matter) as a supplement. Ruminal proportion of acetate tended to decrease and that of propionate was increased (P < 0.05) 2 h after CD‐M dosing. Total viable counts, cellulolytic, sulfate reducing, acetogenic bacteria and protozoa were unaffected while methanogenic bacteria were decreased (P < 0.05) by CD‐M. The plasma concentration of glucose was increased, whereas that of urea‐N was decreased (P < 0.05). Methane was inhibited (P < 0.05) from 36.4 to 30.1 L/kg dry matter intake by the addition of CD‐M. Apparent digestibilities of dry matter and neutral detergent fiber were not affected while that of crude protein was increased (P < 0.05) in the medicated steers. These data suggested that dietary supplementation of CD‐M decreased methane production and improved nutrient use.     

Effects of nitrate addition to a diet on fermentation and microbial populations in the rumen of goats,with special reference to Selenomonas ruminantium having the ability to reduce nitrate and nitrite

This study investigated the effects of dietary nitrate addition on ruminal fermentation characteristics and microbial populations in goats. The involvement of Selenomonas ruminantium in nitrate and nitrite reduction in the rumen was also examined. As the result of nitrate feeding, the total concentration of ruminal volatile fatty acids decreased, whereas the acetate : propionate ratio and the concentrations of ammonia and lactate increased. Populations of methanogens, protozoa and fungi, as estimated by real‐time PCR, were greatly decreased as a result of nitrate inclusion in the diet. There was modest or little impact of nitrate on the populations of prevailing species or genus of bacteria in the rumen, whereas Streptococcus bovis and S. ruminantium significantly increased. Both the activities of nitrate reductase (NaR) and nitrite reductase (NiR) per total mass of ruminal bacteria were increased by nitrate feeding. Quantification of the genes encoding NaR and NiR by real‐time PCR with primers specific for S. ruminantium showed that these genes were increased by feeding nitrate, suggesting that the growth of nitrate‐ and nitrite‐reducing S. ruminantium is stimulated by nitrate addition. Thus, S. ruminantium is likely to play a major role in nitrate and nitrite reduction in the rumen.     

Validation and application of real-time polymerase chain reaction assays for representative rumen bacteria

Real‐time polymerase chain reaction (PCR) assays for 11 representative rumen bacterial species were validated. The sensitivity was tested by using the serially diluted target 16S rDNA from respective bacterial species. The recovery of the target DNA and the assay reproducibility were determined using DNA from rumen fluid spiked with different quantities of the target. Minimum detection levels for the target were 10–100 copies in pure culture. The recovery of the added target ranged from 82.4 to 116.6%. The intra‐ and inter‐assay variations of each assay were <9.4 and <12.6%, respectively. Therefore, the real‐time PCR assays evaluated in the present study are considered to be sufficiently reliable for monitoring all 11 bacterial species in the rumen. The assays were then applied to the monitoring of the bacterial species attached to ruminally incubated rice straw. Among the monitored fibrolytic species, Fibrobacter succinogenes was found to be the most dominant, accounting for 2.61% of total bacteria after 24 h incubation. Selenomonas ruminantium and Streptococcus bovis, non‐fibrolytics, were detected on the rice straw at 8.96% and 1.16% of total bacteria, respectively. Such high levels of non‐fibrolytics on the plant fiber suggest a synergistic relationship between fibrolytics and non‐fibrolytics.     

Feeding value of enset (Ensete ventricosum), Desmodium intortum hay and untreated or urea and calcium oxide treated wheat straw for sheep

Feed intake, in vivo nutrient digestibility and nitrogen utilization were evaluated in male sheep fed different fractions (leaf, pseudostem, corm, whole plant) of enset, untreated or 2% urea‐ and 3% calcium oxide‐ (CaO or lime) treated wheat straw and Desmodium intortum hay as sole diets. All feeds, except D. intortum hay and enset leaf had low crude protein (CP) content. Non‐fiber carbohydrate contents were higher in enset fractions, especially in pseudostem and corm relative to other feeds. Enset leaf and pseudostem had high calcium, phosphorus and manganese contents. Corm, whole enset and D. intortum hay were rich sources of zinc. Daily dry matter and CP intakes were higher (p < 0.05) in sheep fed D. intortum hay (830 and 133 g, respectively) than those fed pseudostem (92 and 7.8 g, respectively). Organic matter digestibilities were highest for corm (0.780) and whole enset (0.776) and lowest for D. intortum hay (0.534) and untreated wheat straw (0.522). The CP digestibility ranged from 0.636 in D. intortum hay to 0.408 in corm. Nitrogen (N) balance was highest (p < 0.05) in D. intortum hay (10.4 g/day) and lowest in corm (?1.3 g/day). Enset leaf could be a useful protein supplement whereas the pseudostem and corm could be good sources of energy.     

Feeding and microbial disorders in horses: 2: Effect of three hay:grain ratios on digesta passage rate and digestibility in ponies

Digesta passage rate and digestibility of diet components (dOM, dNDF and dADF) were measured in the entire digestive tract of six ponies to determine the effects of the dietary hay:grain ratio on the efficiency of fiber digestion. Three diets based on chopped meadow hay and rolled barley were tested in a 3×3 Latin square experiment: 100% hay (100:0, 114gDM/KgBW^0.75/day), 70% hay—30% barley (70:30, 69gDM/KgBW^0.75/day) and 50% hay—50% barley (50:50, 58gDM/KgBW^0.75/day). Particles markers (Europium for hay and Ytterbium for barley) and a solute marker (Cr-EDTA) were used for passage rate measurements. As the proportion of barley in the diet increased, DM intake decreased and mean retention time (MRT) of all markers increased significantly. Even if the barley meal was fed before the hay, excretion rates of all markers were similar. The digestibility of OM increased linearly (P<0.05) whereas that of NDF and ADF decreased (P<0.05). Although the MRTs increased when grain was fed, the efficiency of fiber utilization decreased. Further investigation on fibrolytic microbial ecosystem are required to explain these observations.     

Effects of replacing dietary starch with neutral detergent–soluble fibre on ruminal fermentation,microbial synthesis and populations of ruminal cellulolytic bacteria using the rumen simulation technique (RUSITEC)

A rumen simulation technique (RUSITEC) apparatus with eight 800 ml fermenters was used to investigate the effects of replacing dietary starch with neutral detergent–soluble fibre (NDSF) by inclusion of sugar beet pulp in diets on ruminal fermentation, microbial synthesis and populations of ruminal cellulolytic bacteria. Experimental diets contained 12.7, 16.4, 20.1 or 23.8% NDSF substituted for starch on a dry matter basis. The experiment was conducted over two independent 15‐day incubation periods with the last 8 days used for data collection. There was a tendency that 16.4% NDSF in the diet increased the apparent disappearance of organic matter (OM) and neutral detergent fibre (NDF). Increasing dietary NDSF level increased carboxymethylcellulase and xylanase activity in the solid fraction and apparent disappearance of acid detergent fibre (ADF) but reduced the 16S rDNA copy numbers of Ruminococcus albus in both liquid and solid fractions and R. flavefaciens in the solid fraction. The apparent disappearance of dietary nitrogen (N) was reduced by 29.6% with increased dietary NDSF. Substituting NDSF for starch appeared to increase the ratios of acetate/propionate and methane/volatile fatty acids (VFA) (mol/mol). Replacing dietary starch with NDSF reduced the daily production of ammonia‐N and increased the growth of the solid‐associated microbial pellets (SAM). Total microbial N flow and efficiency of microbial synthesis (EMS), expressed as g microbial N/kg OM fermented, tended to increase with increased dietary NDSF, but the numerical increase did not continue as dietary NDSF exceeded 20.1% of diet DM. Results suggested that substituting NDSF for starch up to 16.4% of diet DM increased digestion of nutrients (except for N) and microbial synthesis, and further increases (from 16.4% to 23.8%) in dietary NDSF did not repress microbial synthesis but did significantly reduce digestion of dietary N.     

Tropical legume supplementation influences microbial protein synthesis and rumen ecology

Four rumen‐fistulated male swamp buffaloes, 5‐year‐old with initiated live weight at 360 ± 12 kg, were randomly assigned according to a 4 × 4 Latin square design to investigate the effect of feeding high level of dried Leucaena leaf (DLL) on feed intake, fermentation efficiency and microbial protein synthesis. The dietary treatments were the feeding levels of DLL at 0, 2, 4 and 6 kg/head/day. All buffaloes were supplemented with concentrate mixtures at 0.1% of body weight, and rice straw was fed ad libitum with the availability of water and mineral block at all time. The results revealed that the total feed intake and nutrient digestibility were significantly improved with the increasing levels of DLL feeding, and the highest was in the buffaloes consuming DLL at 6 kg/head/day. Feeding high levels of DLL did not affect on ruminal pH and temperature, while ammonia nitrogen, blood urea nitrogen and volatile fatty acid concentration were significantly enhanced. Moreover, methane production was dramatically reduced by increasing levels of DLL feeding. Total direct counts of the micro‐organism population were increased with the increasing levels of DLL feeding. According to the application of quantitative PCR to quantity cellulolytic bacteria (16S rRNA) targets, it was found that the population of total bacteria and Fibrobactor succinogenes was affected by treatments, while Ruminococcus flavefaciens and methanogen population were significantly decreased as buffaloes were fed with DLL. The nitrogen balance and microbial nitrogen supply were remarkably improved with the increasing levels of DLL feeding. Based on this study, it could be concluded that high levels of DLL feeding at 6 kg/head/day could enhance feed intake, nutrient digestibility, rumen fermentation efficiency and microbial protein synthesis in swamp buffaloes fed on rice straw without any adverse effect.