Ruminobacter Amylophilus Research Articles

Application of the bacterial strains Ruminobacter amylophilus, Fibrobacter succinogenes and Enterococcus faecium for growth promotion in maize and soybean plants

Ruminobacter amylophilus, Fibrobacter succinogenes and Enterococcus faecium have characteristics that are similar to those of plant growth-promoting bacteria and can be used to promote plant development and reduce production costs. These bacteria were isolated from fistulated ruminants and are gram-negative, anaerobic or facultative anaerobic. These bacteria are frequently used to increase animal productivity through the production of many enzymes responsible for the carbon cycle and the release of other nutrients by organic matter decomposition. The bacteria R. amylophilus, F. succinogenes and E. faecium have growth promotion abilities, such as phosphorus solubilization, nitrogen promotion, and indole acetic acid and siderophore production. Tests were performed under greenhouse conditions with soybean and maize crops with five treatments and six replications. The first treatment was the control (without inoculant); the other treatments included each species of bacteria, and there was a treatment with a mixture (mix) of the three bacteria. F. succinogenes increased the root dry mass of maize by 21.4%, as well as the nitrogen and phosphorus contents, compared to the control. R. amylophilus and E. faecium decreased the phosphorus concentration in shoots of maize, and R. amylophilus increased the soil biomass carbon by 76.39% compared to the mix under maize cultivation, while E. faecium decreased the soil biomass carbon by 56.78% compared to the mix under soybean cultivation. The present study verified that Ruminobacter amylophilus, Fibrobacter succinogenes and Enterococcus faecium presented plant growth-related abilities and could be used to improve plant development, reducing the necessity of chemical fertilizers

Effects of dietary laccase supplementation on growth performance, nutrient digestion, rumen fermentation and microbiota in dairy bulls

This study investigated the impacts of dietary laccase addition on performance, nutrient digestion, rumen fermentation and microbiota in bulls. Eight Holstein bulls with rumen cannula were allocated in a repeated 4 × 4 Latin square experiment design into four groups: control, low-laccase (LL), medium-laccase (ML) and high-laccase (HL) with 0, 0.2, 0.4 and 0.6 g/kg dietary dry matter (DM) of laccase addition, respectively. Addition of laccase did not influence DM intake but higher average daily gain and lower feed conversion ratio were found for bulls in ML group. Apparent nutrients digestibility elevated linearly with increasing laccase supplementation. Digestibility of DM and organic matter in ML, crude protein (CP) in ML and HL as well as neutral detergent fibre (NDF) and acid detergent fibre in LL, ML and HL were higher. Increasing laccase addition quadratically decreased ruminal pH, quadratically increased total volatile fatty acids content and changed ruminal fermentation mode to more propionate formation. Ruminal ammonia-N concentration decreased linearly. Ruminal degradability of concentrate CP and corn silage DM were higher for bulls in ML, but degradability of corn silage NDF was higher for bulls in ML and HL groups. Activities of cellobiase, α-amylase and laccase were higher for bulls in ML group. Populations of total bacteria, methanogens, Fibrobacter succinogenes, Ruminococcus albus, Butyrivibrio fibrisolvens, Prevotella ruminicola and Ruminobacter amylophilus increased linearly, but fungi and R. flavefaciens increased quadratically. Urine excretion of total purine derivatives was higher for bulls receiving medium or high level of laccase supplementation. The results suggested that dietary laccase supplementation increased performance and nutrient digestibility, and the appropriate addition level of laccase was 0.4 g/kg DM in bulls.

Dietary Energy Levels Affect Rumen Bacterial Populations that Influence the Intramuscular Fat Fatty Acids of Fattening Yaks (Bos grunniens).

Simple SummaryYak, a bovid animal, is the predominant livestock on the Qinghai–Tibet Plateau. Rumen is an important digestive organ for ruminants, such as cattle, yak, and sheep. Rumen bacteria play a crucial role in dietary energy digestion of yaks and in their adaptation to the plateau environment. Dietary energy levels affect rumen bacterial populations and lipid deposition in the meat of ruminants. The intramuscular fat fatty acid profile is important for meat quality and human health. This study was conducted to determine the rumen bacterial populations affected by dietary energy levels and understand their relationship with intramuscular fat fatty acids. The results found that increasing dietary energy significantly increased ruminal propionate concentration and reduced the ammonia concentration. High dietary energy increased the ratio of Firmicutes to Bacteroidetes and mainly increased ruminal amylolytic and propionate-producing bacteria populations. Ruminal acetate and propionate were positively related to intramuscular saturated fatty acid content, and Prevotella was positively related to intramuscular polyunsaturated fatty acid content and negatively related to intramuscular saturated fatty acid content. This study gives insights into how the effects of dietary energy on rumen bacterial populations are related to intramuscular fat fatty acids of yaks.The yak rumen microflora has more efficient fiber-degrading and energy-harvesting abilities than that of low-altitude cattle; however, few studies have investigated the effects of dietary energy levels on the rumen bacterial populations and the relationship between rumen bacteria and the intramuscular fatty acid profile of fattening yaks. In this study, thirty yaks were randomly assigned to three groups. Each group received one of the three isonitrogenous diets with low (3.72 MJ/kg), medium (4.52 MJ/kg), and high (5.32 MJ/kg) levels of net energy for maintenance and fattening. After 120 days of feeding, results showed that increasing dietary energy significantly increased ruminal propionate fermentation and reduced ammonia concentration. The 16S rDNA sequencing results showed that increasing dietary energy significantly increased the ratio of Firmicutes to Bacteroidetes and stimulated the relative abundance of Succiniclasticum, Saccharofermentans, Ruminococcus, and Blautia populations. The quantitative real-time PCR analysis showed that high dietary energy increased the abundances of Streptococcus bovis, Prevotella ruminicola, and Ruminobacter amylophilus at the species level. Association analysis showed that ruminal acetate was positively related to some intramuscular saturated fatty acid (SFA) contents, and Prevotella was significantly positively related to intramuscular total polyunsaturated fatty acid content and negatively related to intramuscular total SFA content. This study showed that high dietary energy mainly increased ruminal amylolytic and propionate-producing bacteria populations, which gave insights into how the effects of dietary energy on rumen bacteria are related to intramuscular fat fatty acids of fattening yaks.

Effects of copper sulphate and coated copper sulphate addition on lactation performance, nutrient digestibility, ruminal fermentation and blood metabolites in dairy cows.

Coated copper sulphate (CCS) could be used as a Cu supplement in cows. To investigate the influences of copper sulphate (CS) and CCS on milk performance, nutrient digestion and rumen fermentation, fifty Holstein dairy cows were arranged in a randomised block design to five groups: control, CS addition (7·5 mg Cu/kg DM from CS) or CCS addition (5, 7·5 and 10 mg Cu/kg DM from CCS, respectively). When comparing Cu source at equal inclusion rates (7·5 mg/kg DM), cows receiving CCS addition had higher yields of fat-corrected milk, milk fat and protein; digestibility of DM, organic matter (OM) and neutral-detergent fibre (NDF); ruminal total volatile fatty acid (VFA) concentration; activities of carboxymethyl cellulase, cellobiase, pectinase and α-amylase; populations of Ruminococcus albus, Ruminococcus flavefaciens and Fibrobacter succinogenes; and liver Cu content than cows receiving CS addition. Increasing CCS addition, DM intake was unchanged, yields of milk, milk fat and protein; feed efficiency; digestibility of DM, OM, NDF and acid-detergent fibre; ruminal total VFA concentration; acetate:propionate ratio; activity of cellulolytic enzyme; populations of total bacteria, protozoa and dominant cellulolytic bacteria; and concentrations of Cu in serum and liver increased linearly, but ruminal propionate percentage, ammonia-N concentration, α-amylase activity and populations of Prevotella ruminicola and Ruminobacter amylophilus decreased linearly. The results indicated that supplement of CS could be substituted with CCS and addition of CCS improved milk performance and nutrient digestion in dairy cows.

Effects of betaine and rumen-protected folic acid supplementation on lactation performance, nutrient digestion, rumen fermentation and blood metabolites in dairy cows

This study evaluated the effects of betaine (BT) and rumen-protected folic acid (RPFA) on lactation performance, nutrient digestion, rumen fermentation and blood metabolites in dairy cows. Thirty-six mid-lactation Holstein dairy cows were assigned to one of four treatments in a 2 × 2 factorial randomized block design. Supplemental RPFA (0 [RPFA-] or 5.2 mg/kg dietary dry matter [DM] of folic acid [FA] from RPFA [RPFA+]) and BT (0 [BT-] or 4.0 g/kg DM of BT [BT+]) was mixed into the daily ration, respectively. The experiment lasted for 105 days with the first 15 days for adaptation and the following 90 days for sample collection. The RPFA × BT interaction was significant for blood folate concentration which decreased with BT addition in RPFA- diet but increased with BT addition in RPFA + diet. Dry matter intake and body weight change were similar among treatments. Dietary RPFA or BT addition increased yield of milk and milk fat but higher milk protein yield was only observed for RPFA addition. Feed efficiency was increased due to RPFA or BT addition. Apparent total-tract digestibility of DM, organic matter, crude protein, neutral detergent fibre and acid detergent fibre increased with RPFA or BT addition. Ruminal pH decreased with RPFA addition but was unchanged with BT addition. Dietary RPFA or BT addition increased ruminal total volatile fatty acids concentration and altered rumen fermentation mode to more acetate production. Ammonia-N concentration was unchanged with RPFA addition but decreased with BT addition. Addition of RPFA or BT increased activity of carboxymethyl-cellulase, xylanase and protease as well as population of total bacteria, protozoa, Ruminococcus (R.) albus and R. flavefaciens. Population of Prevotella ruminicola was higher for RPFA addition but activity of cellobiase and population of total fungi and Ruminobacter amylophilus was higher for BT addition. Blood concentrations of glucose, non-esterified fatty acid, beta-hydroxybutyrate and homocysteine were similar among treatments. Blood total protein and albumin concentrations were increased with RPFA addition but were not affected by BT addition. The data indicated that addition of RPFA or BT had beneficial impacts on lactation performance, nutrient digestion and ruminal fermentation and BT addition might increase FA utilization efficiency in dairy cows receiving diet without RPFA.

Effects of rumen-protected folic acid and betaine supplementation on growth performance, nutrient digestion, rumen fermentation and blood metabolites in Angus bulls.

This study evaluated the effects of rumen-protected folic acid (RPFA) and betaine (BT) on growth performance, nutrient digestion and blood metabolites in bulls. Forty-eight Angus bulls were blocked by body weight and randomly assigned to four treatments in a 2 × 2 factorial design. BT of 0 or 0·6 g/kg DM was supplemented to diet without or with the addition of 6 mg/kg DM of folic acid from RPFA, respectively. Average daily gain increased by 25·2 and 6·29 % for addition of BT without RPFA and with RPFA, respectively. Digestibility and ruminal total volatile fatty acids of neutral-detergent fibre and acid-detergent fibre increased, feed conversion ratio and blood folate decreased with the addition of BT without RPFA, but these parameters were unchanged with BT addition in diet with RPFA. Digestibility of DM, organic matter and crude protein as well as acetate:propionate ratio increased with RPFA or BT addition. Ruminal ammonia-N decreased with RPFA addition. Activity of carboxymethyl cellulase, cellobiase, xylanase, pectinase and protease as well as population of total bacteria, protozoa, Fibrobacter succinogenes and Ruminobacter amylophilus increased with RPFA or BT addition. Laccase activity and total fungi, Ruminococcus flavefaciens and Prevotella ruminicola population increased with RPFA addition, whereas Ruminococcus albus population increased with BT addition. Blood glucose, total protein, albumin, growth hormone and insulin-like growth factor-1 increased with RPFA addition. Addition of RPFA or BT decreased blood homocysteine. The results indicated that addition of BT stimulated growth and nutrient digestion in bulls only when RPFA was not supplemented.

Invitro rumen fermentation of feed substrates added with chestnut tannins or an extract from Stevia rebaudiana Bertoni.

Rumen fermentation parameters and microbiota were evaluated in 3 in vitro rumen fermentation experiments after addition of chestnut tannins (CWT) or an extract from Stevia rebaudiana Bertoni (SB) to substrates. A control (CTR) substrate was fermented alone or added with 1.5% of CWT or SB extracts in a batch culture system (Exp. 1, fermentation in 500 mL for 24 h) and in a subsequent continuous culture system (Exp. 2, fermentation in 2 L bottles for 9 d). Experiment 3 used the fermentation system of Exp. 1 and tested 7 doses of each extract added to CTR (additions of 0.2%, 0.4%, 0.6%, 0.8%, 1.0%, 1.2% and 1.4% for 48 h). The addition of CWT lowered (P < 0.01) the in vitro rumen ammonia concentration in all experiments and reduced the protozoa counts in Exp. 1 (P < 0.05). In contrast, the SB extract did not modify the ammonia concentrations, but significantly lowered the protozoa counts in all 3 experiments (reduction of 47% and 20% in Exp. 1 and 2, P < 0.05; and a quadratic reduction in Exp. 3, R2 = 0.63, P < 0.01). Neither extract affected the fermentation in terms of gas production (Exp. 1 and 3) nor volatile fatty acids (VFA) yield (Exp. 1 and 2), if we exclude a reduction at the highest CWT concentration in Exp. 3. Changes in VFA profile were induced by CWT and were limited to reductions in the iso-valerate (P < 0.01, in Exp. 2) and iso-butyrate levels (P < 0.01, Exp. 2). The CWT increased the abundance of Prevotella ruminicola and Selenomonas ruminantium and decreased that of Ruminobacter amylophilus (P < 0.01, P < 0.05 and P < 0.05, respectively). The SB extract increased the relative abundance of Treponema saccarophylum (P < 0.05). Both of the studied substances had an impact on rumen metabolism, with SB reducing protozoa counts and CWT lowering the rumen ammonia concentration. The effects of both extracts on the rumen were appreciable at low dietary doses, and the negative impacts on fermentation were limited to the reduction in protein degradation with the addition of CWT.

Effects of guanidinoacetic acid supplementation on growth performance, nutrient digestion, rumen fermentation and blood metabolites in Angus bulls

Guanidinoacetic acid (GAA) can improve the growth performance of bulls. This study investigated the influences of GAA addition on growth, nutrient digestion, ruminal fermentation and serum metabolites in bulls. Forty-eight Angus bulls were randomly allocated to experimental treatments, that is, control, low-GAA (LGAA), medium-GAA (MGAA) and high-GAA (HGAA), with GAA supplementation at 0, 0.3, 0.6 and 0.9 g/kg DM, respectively. Bulls were fed a basal diet containing 500 g/kg DM concentrate and 500 g/kg DM roughage. The experimental period was 104 days, with 14 days for adaptation and 90 days for data collection. Bulls in the MGAA and HGAA groups had higher DM intake and average daily gain than bulls in the LGAA and control groups. The feed conversion ratio was lowest in MGAA and highest in the control. Bulls receiving 0.9 g/kg DM GAA addition had higher digestibility of DM, organic matter, NDF and ADF than bulls in other groups. The digestibility of CP was higher for HGAA than for LGAA and control. The ruminal pH was lower for MGAA, and the total volatile fatty acid concentration was greater for MGAA and HGAA than for the control. The acetate proportion and acetate-to-propionate ratio were lower for MGAA than for LGAA and control. The propionate proportion was higher for MGAA than for control. Bulls receiving GAA addition showed decreased ruminal ammonia N. Bulls in MGAA and HGAA had higher cellobiase, pectinase and protease activities and Butyrivibrio fibrisolvens, Prevotella ruminicola and Ruminobacter amylophilus populations than bulls in LGAA and control. However, the total protozoan population was lower for MGAA and HGAA than for LGAA and control. The total bacterial and Ruminococcus flavefaciens populations increased with GAA addition. The blood level of creatine was higher for HGAA, and the activity of l-arginine glycine amidine transferase was lower for MGAA and HGAA, than for control. The blood activity of guanidine acetate N-methyltransferase and the level of folate decreased in the GAA addition groups. The results indicated that dietary addition of 0.6 or 0.9 g/kg DM GAA improved growth performance, nutrient digestion and ruminal fermentation in bulls.

PSIX-14 Effects of substituting dietary corn with beet pulp on growth performance and microbial profiles in the rumen of Korean cattle steers

Abstract This experiment evaluated the effects of substituting corn with beet pulp (BP) on growth performance and ruminal fermentation characteristics and microbiomes in fattening steers. Twelve Korean cattle steers (body weight, 485 ± 19 kg; age, 18.0 ± 0.17 months) were divided into corn and BP groups. Both groups were fed the basal concentrate to meet 80% of the nutrient requirements. For the corn group, corn (dry matter) was supplemented for the remaining 20% of requirement by top dressing; the BP group was supplemented with 14.88% BP plus 1.92% fat to provide the iso-energy content as the corn group. Steers were fed oat grass (2 kg as-fed basis) daily. Body weight was measured, and rumen fluid was collected after 3 h of feeding using oral stomach tube methods: at the start of the experiment then at 4-week intervals. Feed intake, average daily gain, and gain-to-feed ratio were not (P &amp;gt; 0.05) affected by BP substitution; nor were acetate, propionate, butyrate, or total volatile fatty acid concentrations in the rumen fluid (P &amp;gt; 0.05). BP substitution increased (P &amp;lt; 0.05) the acetate to propionate ratio at 4, 8, and 12 weeks. Genomic DNA was extracted from pellets of rumen contents, and microbial populations were measured using real-time PCR at 8 and 12 weeks. BP substitution increased relative abundances of cellulolytic bacteria including Fibrobacter succinogenes (P = 0.01) and Ruminococcus albus (P = 0.04) at 12 weeks, whereas it did not affect amylolytic bacteria, including Succinimonas amylolytica, Ruminobacter amylophilus, and Streptococcus bovis. At 8 weeks, none of the microbial populations were altered by BP substitution. In conclusion, BP could provide a good energy source substitute for corn without affecting growth performance in the fattening period of beef cattle. Increased relative abundances of several cellulolytic bacteria supported the increased ruminal acetate to propionate ratio.

Effects of folic acid and sodium selenite on growth performance, nutrient digestion, ruminal fermentation and urinary excretion of purine derivatives in Holstein dairy calves

This study evaluated the effects of folic acid (FA) and sodium selenite (SS) supplements on growth performance, nutrient digestion, ruminal fermentation, microbial enzyme activity, microflora and urinary excretion of purine derivatives (PD) in dairy calves. Thirty-six Holstein calves (63 ± 3.4 days of age and 83 ± 2.1 kg of body weight) were randomly assigned to four treatments in a 2 × 2 factorial arrangement. Two levels of FA (0 or 3.0 mg/kg of FA) and SS (0 or 0.3 mg/kg of Se from SS) were added into diets on a dry matter (DM) basis. Nutrient intake and average daily gain increased with FA addition, but were not affected by SS addition. Both supplements increased total tract digestibility of DM, organic matter and neutral detergent fiber. Ruminal pH and ammonia N concentration decreased, but total volatile fatty acids concentration increased with FA or SS addition. Acetate molar proportion and acetate to propionate ratio increased, while propionate molar proportion decreased with FA addition. The unchanged acetate molar proportion, higher propionate molar proportion and lower acetate to propionate ratio were observed with SS addition compared with the control. Activities of xylanase, pectinase, α-amylase and protease as well as populations of total bacteria, protozoa and methanogens were higher for both supplements compared with the control. Carboxymethyl-cellulase activity and Prevotella ruminicola population increased with FA addition. The Ruminobacter amylophilus population decreased with SS addition. The FA×SS interaction was significant for total tract digestibility of crude protein and acid detergent fiber, populations of total fungi, Ruminococcus (R.) albus, R. flavefaciens, Fibrobacter succinogenes and Butyrivibrio fibrisolvens as well as urinary total PD excretion; these variables increased with SS addition and the increase was greater when the diet was not supplemented with FA than with FA supplemented diet. The results indicated that dietary FA and/or SS addition stimulated nutrient digestion, ruminal fermentation, microbial growth and urinary total PD excretion.

Effects of soybean oil and dietary copper levels on nutrient digestion, ruminal fermentation, enzyme activity, microflora and microbial protein synthesis in dairy bulls

ABSTRACT The study evaluated the effects of soybean oil (SO) and dietary copper levels on nutrient digestion, ruminal fermentation, enzyme activity, microflora and microbial protein synthesis in dairy bulls. Eight Holstein rumen-cannulated bulls (14 ± 0.2 months of age and 326 ± 8.9 kg of body weight) were allocated into a replicated 4 × 4 Latin square design in a 2 × 2 factorial arrangement with factors being 0 or 40 g/kg dietary dry matter (DM) of SO and 0 or 7.68 mg/kg DM of Cu from copper sulphate (CS). The basal diet contained per kg DM 500 g of corn silage, 500 g of concentrate, 28 g of ether extract (EE) and 7.5 mg of Cu. The SO × CS interaction was significant (p < 0.05) for ruminal propionate proportion and acetate to propionate ratio. Dietary SO addition increased (p < 0.05) intake and total tract digestibility of EE but did not affect average daily gain (ADG) of bulls. Dietary CS addition did not affect nutrient intake but increased (p < 0.05) ADG and total tract digestibility of DM, organic matter, crude protein and neutral detergent fibre. Ruminal pH was not affected by treatments. Dietary SO addition did not affect ruminal total volatile fatty acids (VFA) concentration, decreased (p < 0.05) acetate proportion and ammonia N and increased (p < 0.05) propionate proportion. Dietary CS addition did not affect ammonia N, increased (p < 0.05) total VFA concentration and acetate proportion and decreased (p < 0.05) propionate proportion. Acetate to propionate ratio decreased (p < 0.05) with SO addition and increased (p < 0.05) with CS addition. Dietary SO addition decreased (p < 0.05) activity of carboxymethyl cellulase, cellobiase and xylanase as well as population of fungi, protozoa, methanogens, Ruminococcus albus and R. flavefaciens but increased (p < 0.05) α-amylase activity and population of Prevotella ruminicola and Ruminobacter amylophilus. Dietary CS addition increased (p < 0.05) activity of cellulolytic enzyme and protease as well as population of total bacteria, fungi, protozoa, methanogens, primary cellulolytic and proteolytic bacteria. Microbial protein synthesis was unchanged with SO addition but increased (p < 0.05) with CS addition. The results indicated that the addition of CS promoted nutrient digestion and ruminal fermentation by stimulating microbial growth and enzyme activity but did not relieve the negative effects of SO addition on ruminal fermentation in dairy bulls.

Effects of pantothenic acid and folic acid supplementation on total tract digestibility coefficient, ruminal fermentation, microbial enzyme activity, microflora and urinary purine derivatives in dairy bulls

AbstractThe effects of pantothenic acid (PA) and folic acid (FA) addition on digestibility coefficient, ruminal fermentation and urinary purine derivative (PD) excretion in dairy bulls were evaluated. Eight rumen-cannulated Holstein dairy bulls were allocated to a replicated 4 × 4 Latin square design according to a 2 × 2 factorial arrangement. Diets were supplemented with two levels of FA (0 or 8.0 mg/kg dietary dry matter [DM]) and two of PA (0 or 60 mg/kg DM). The PA × FA interaction was not significant for all variables. Both supplements increased DM intake and average daily gain, but decreased a feed conversion ratio. Digestibility of DM, organic matter, crude protein and neutral detergent fibre increased, but ether extract digestibility was unchanged for both supplements. Digestibility of acid detergent fibre only increased with FA supplementation. For both supplements, ruminal pH and ammonia nitrogen (N) decreased, but total volatile fatty acid (VFA) concentration increased. Acetate proportion only increased with FA supplementation. Propionate proportion decreased for both supplements. Consequently, the acetate to propionate ratio increased. For both supplements, activity of xylanase and pectinase, population of Ruminococcus albus, R. flavefaciens, Fibrobacter succinogenes and Ruminobacter amylophilus and total PD excretion increased. Additionally, activity of carboxymethylcellulase, cellobiase, α-amylase and protease, and population of total bacteria, fungi, protozoa, methanogens, Butyrivibrio fibrisolvens and Prevotella ruminicola increased with FA addition. The results suggested that PA and FA supplementation stimulated ruminal microbial growth and enzyme activity, resulting in an increased digestibility coefficient and ruminal total VFA concentration in dairy bulls.

Effects of replacing alfalfa hay with Moringa oleifera leaves and peduncles on intake, digestibility, and rumen fermentation in dairy cows

To study the effects of replacing alfalfa hay with Moringa oleifera leaves and peduncles in diets on intake, digestibility, and rumen fermentation in dairy cows, eight multiparous Holsteins cows with similar DIM (day in milk, 93 ± 18 d) and body weights (624 ± 38 kg) were used in a crossover design with two dietary treatments. The treatment diets contained similar concentrate mixtures and corn silage but different proportions of M. oleifera leaves and peduncles. The treatments were a (1) control diet (0% M. oleifera feed) and (2) M. oleifera diet (10.85% M. oleifera feed). The M. oleifera feed used M. oleifera leaves and M. oleifera peduncles at a ratio of 3:2 and replaced portions of alfalfa hay. The experimental period was 18 d, with 15 d for treatment adaptation and 3 d for data collection. Compared with the control diet, the dry matter intake and apparent digestibility of dry matter, crude protein, acid detergent fiber and neutral detergent fiber in the M. oleifera diet were significantly increased. No difference was observed in ruminal pH; however, the ammonia-N concentrate was lower for the M. oleifera diet than the control diet. The concentrations of total ruminal volatile fatty acids, acetate, propionate, and butyrate were higher in cows fed the M. oleifera diet than in those fed the control diet. The results of ruminal microbes showed that those consuming the M. oleifera diet had greater relative expression level of Fibrobacter succinogenes, Butyrivibrio fibrisolvens, Ruminobacter amylophilus, Streptococcus bovis, Prevotella ruminicola which were observed for the control diet than the M. oleifera diet. These results show that the M. oleifera diet can improve intake and apparent digestibility of nutrients, and rumen fermentation in dairy cows.

Effects of short-term fasting on in vivo rumen microbiota and in vitro rumen fermentation characteristics.

ObjectiveFasting may lead to changes in the microbiota and activity in the rumen. In the present study, the effects of fasting on rumen microbiota and the impact of fasting on in vitro rumen fermentation were evaluated using molecular culture-independent methods.MethodsThree ruminally cannulated Holstein steers were fed rice straw and concentrates. The ruminal fluids were obtained from the same steers 2 h after the morning feeding (control) and 24 h after fasting (fasting). The ruminal fluid was filtrated through four layers of muslin, collected for a culture-independent microbial analysis, and used to determine the in vitro rumen fermentation characteristics. Total DNA was extracted from both control and fasting ruminal fluids. The rumen microbiota was assessed using denaturing gradient gel electrophoresis (DGGE) and quantitative polymerase chain reaction. Microbial activity was evaluated in control and fasting steers at various intervals using in vitro batch culture with rice straw and concentrate at a ratio of 60:40.ResultsFasting for 24 h slightly affected the microbiota structure in the rumen as determined by DGGE. Additionally, several microorganisms, including Anaerovibrio lipolytica, Eubacterium ruminantium, Prevotella albensis, Prevotella ruminicola, and Ruminobacter amylophilus, decreased in number after fasting. In addition, using the ruminal fluid as the inoculum after 24 h of fasting, the fermentation characteristics differed from those obtained using non-fasted ruminal fluid. Compared with the control, the fasting showed higher total gas production, ammonia, and microbial protein production (p<0.05). No significant differences, however, was observed in pH and dry matter digestibility.ConclusionWhen in vitro techniques are used to evaluate feed, the use of the ruminal fluid from fasted animals should be used with caution.

Effects of concentrate-to-forage ratios and 2-methylbutyrate supplementation on ruminal fermentation, bacteria abundance and urinary excretion of purine derivatives in Chinese Simmental steers.

This study evaluated the effects of dietary concentrate levels and 2-methylbutyrate (2MB) supplementation on performance, ruminal fermentation, bacteria abundance, microbial enzyme activity and urinary excretion of purine derivatives (PD) in steers. Eight ruminally cannulated Simmental steers (12months of age; 389±3.7kg of body weight) were used in a replicated 4×4 Latin square design with a 2×2 factorial arrangement. Moderate-concentrate (400g/kg diet [MC]) or high-concentrate (600g/kg diet [HC]) diets were fed with or without 2MB (0g/day [2MB-] or 15.0g/day [2MB+]). Dry matter intake and average daily gain increased, but feed conversion ratio decreased with the HC diet or 2MB supplementation. Ruminal pH decreased, but total volatile fatty acid increased with the HC diet or 2MB supplementation. Molar proportion of acetate and acetate-to-propionate ratio decreased with the HC diet, but increased with 2MB supplementation. Propionate molar proportion and ruminal NH3 -N content increased with the HC diet, but decreased with 2MB supplementation. Neutral detergent fibre degradability decreased with the HC diet, but increased with 2MB supplementation. Crude protein degradability increased with the HC diet or 2MB supplementation. Abundance of Ruminococcus albus, Ruminococcus flavefaciens, Fibrobacter succinogenes and Bufyrivibrio fibrisolvens as well as activities of carboxymethyl cellulase, cellobiase, xylanase and pectinase decreased with the HC diet, but increased with 2MB supplementation. However, abundance of Prevotella ruminicola and Ruminobacter amylophilus as well as activities of α-amylase and protease increased with the HC diet or 2MB supplementation. Total PD excretion also increased with the HC diet or 2MB supplementation. The results suggested that growth performance, ruminal fermentation, CP degradability and total PD excretion increased with increasing dietary concentrate level from 40% to 60% or 2MB supplementation. The observed diet×2MB interaction indicated that supplementation of 2MB was more efficacious for improving growth performance, ruminal fermentation and total PD excretion with promoted ruminal bacteria abundance and enzyme activity in the MC diet than in the HC diet.

Effects of certain long-chain fatty acid combinations on the ruminal microbe species relating to fermentation type in vitro

This experiment was conducted to investigate the effects of certain long-chain fatty acid combinations on ruminal microbial community in vitro. Three cows fitted with permanent ruminal cannulas were used to provide rumen liquor for the in vitro trail. The treatments were fatty acid combinations of stearic acid, oleic acid, linoleic acid and linolenic acid, which respectively were group A (calcium palmitate, control), B (1.5%, 1.0%, 0.5% and 1.5%, acetic type fermentation), C (1.5%, 1.0%, 1.5% and 1.0%, propionic acid type fermentation), D (1.0%, 1.5%, 1.5% and 0.5%, butyric acid type fermentation). Three goats fitted with rumen cannula were used to provide the culture medium was collected for the measurement of the rumen microorganism in vitro culture. Culture medium was collected at 0, 3, 6, 12, 18 and 24 h. The results demonstrated that, except for Ruminococcus albus, all the other bacterial genus had significant differences between groups (P less than 0.05). The Fibrobacter succinogenes and Ruminococcusflavefaciens, Clostridium proteoclasticum and Ruminobacter amylophilus were higher in group B; Butyrivibrio fibrisolvens, Megasphaera elsdenii and Ruminococcus bromii were higher in group C; while Pseudobutyrivibrio ruminis was higher in group D. Different higher species in different treatments described above related to their fermentation type. It was also observed that, bacteria were the dominant flora in rumen and Megasphaera elsdeniiwere the dominant species in bacterial fraction no matter what the treatment is. It was therefore concluded that, certain fatty acid combinations have remarkable effects on the amount of ruminal specific bacteria species,mainly related to their fermentation type.

Dietary supplements during the cold season increase rumen microbial abundance and improve rumen epithelium development in Tibetan sheep.

Livestock on the Qinghai-Tibetan Plateau are faced with extreme harsh winters and are often in negative energy balance during this period. Dietary supplementation can improve growth performance of Tibetan sheep and, consequently, we hypothesized that it would also increase microbial abundance and rumen epithelium development. To test this hypothesis, we examined the effect of feed supplementation during the cold season on rumen microbes, fermentation, epithelium development, and absorptive capability in Tibetan sheep. Eighteen 1-yr-old ewes (BW = 29.4 ± 1.79, kg) were offered oat hay ad libitum for 60 d and divided randomly into three groups: 1) no supplement; control group (CON); 2) urea-molasses lick block supplement (BS); and 3) concentrate feed supplement (CS). The ADG of CS ewes (143.3, g/d) was greater (P < 0.05) than BS ewes (87.9, g/d), which was greater (P < 0.05) than CON ewes (44.5, g/d). Serum concentrations of GH, IGF-1, and IGF-2 in the CS and BS groups were greater than in the CON group (P < 0.05). Greater relative abundance of protozoa, Ruminococcus albus, Fibrobacter succinogenes, Streptococcus bovis, and Ruminobacter amylophilus was observed in the CS and BS groups than in the CON group (P < 0.05), and relative abundances of rumen fungi, Butyrivibrio fibrisolvens, and Prevotella ruminicola in the CS group were greater than in the BS and CON groups (P < 0.05). Ruminal total VFA, ammonia, and microbial protein concentrations in the CS and BS groups were greater than in the CON group (P < 0.05), and in the CS group were greater than in the BS group (P < 0.05). Ruminal papillae width and surface area in the CS and BS groups were greater than in the CON group (P < 0.05), while in the CS group were greater than in the BS group (P < 0.05). The mRNA expressions of IGFBP5, NHE1 (sodium/hydrogen antiporter, isoform 1), DRA (downregulated in adenoma), and Na+/K+-ATPase (sodium/potassium ATPase pump) in ruminal epithelium were greater in the CS and BS groups than in the CON group (P < 0.05), and in the CS group was greater than in the BS group (P < 0.05), while NHE3 (sodium/hydrogen antiporter, isoform 3), MCT1 (monocarboxylate transporter 1), and MCT4 (monocarboxylate transporter 4) mRNA expressions in the CS group were greater than in the BS and CON groups (P < 0.05). It was concluded that supplementing Tibetan sheep during the cold season increases rumen microbial abundance and improves fermentation parameters, rumen epithelium development, and absorptive capability.

Dietary Alfalfa and Calcium Salts of Long-Chain Fatty Acids Alter Protein Utilization, Microbial Populations, and Plasma Fatty Acid Profile in Holstein Freemartin Heifers.

This study presented the effects of alfalfa and calcium salts of long-chain fatty acids (CSFA) on feed intake, apparent digestibility, rumen fermentation, microbial community, plasma biochemical parameters, and fatty acid profile in Holstein freemartin heifers. Eight Holstein freemartin heifers were randomly divided into a 4 × 4 Latin Square experiment with 2 × 2 factorial diets, with or without alfalfa or CSFA. Dietary supplementation of CSFA significantly increased the apparent digestibility of dry matter, crude protein, neutral detergent fiber, organic matter, and significantly reduced N retention (P < 0.05). CSFA increased the concentration of ammonia nitrogen in the ruminal fluid (P < 0.05), but alfalfa increased the concentration of valerate and isovalerate (P < 0.05). CSFA increased the concentration of ammonia nitrogen and the relative population of Streptococcus bovis in the rumen (P < 0.05) and inhibited the relative population of Ruminococcus flavefaciens, methanogens, and protozoa (P < 0.05). Alfalfa instead of Leymus chinensis increased the relative population of Butyrivibrio fibrisolvens and Ruminobacter amylophilus in the rumen (P < 0.05) and reduced the relative population of the Ruminococcus albus and Megasphaera elsdenii (P < 0.05). Supplemental CSFA increased the concentration of cholesterol and low-density lipoprotein cholesterol in the plasma (P < 0.05). And it also altered the composition of fatty acids in the plasma, which was expressed in reducing saturated fatty acid (ΣSFA) ratio and C14-C17 fatty acids proportion except C16:0 (P < 0.05) and increasing the proportion of polyunsaturated fatty acid (ΣPUFA) and unsaturated fatty acid (ΣUFA) (P < 0.05). The results showed that alfalfa and CSFA had interaction effect on the apparent digestibility of ether extracts, plasma triglyceride concentration, isobutyrate concentration, and Ruminococcus albus relative abundance in the rumen. It was concluded that alfalfa substituting Leymus chinensis did not change the apparent digestibility of nutrients in the final stage of fattening Holstein freemartin heifers, while CSFA increased the cholesterol and the proportion of unsaturated fatty acids in plasma. Alfalfa and CSFA had mutual interaction effect on fat digestion and plasma triglycerides.

Effects of dietary protein levels and rumen-protected pantothenate on ruminal fermentation, microbial enzyme activity and bacteria population in Blonde d'Aquitaine × Simmental beef steers

This study was undertaken to investigate the effects of dietary crude protein (CP) levels and rumen-protected pantothenate (RPP) supplementation on ruminal digestion kinetics, fermentation, microbial enzymatic activity, and selected bacterial species relative abundance in beef steers. Eight ruminally cannulated crossbred beef steers, with 15 months of age and 462±9kg of body weight (BW), were used in a duplicated 4×4 Latin square design by a 2×2 factorial arrangement. Low protein (113.7g/kg of CP [LP] or high protein (133.9g/kg of CP [HP]) diets were fed with or without RPP supplementation (0g [RP-] or 0.48g per kg dietary dry matter (DM) [RP+]). Steers were fed a total mixed ration containing dietary concentrate to corn silage ratio of 50:50 on a DM basis. There were no RPP×CP interactions for ruminal digestion kinetics, fermentation, microbial enzymatic activity, and selected bacterial species relative abundance, except for Ruminobacter amylophilus. Mean ruminal pH decreased with RPP supplementation, but was unaffected by dietary CP levels. Ruminal total VFA concentration increased with increasing dietary CP levels or RPP supplementation. Ruminal acetate percentage was similar among treatments, whereas propionate percentage decreased with increasing dietary CP levels or RPP supplementation. Consequently, the acetate to propionate ratio increased with increasing dietary CP levels or RPP supplementation. Ruminal ammonia-N content was unchanged among treatments. Both in situ ruminal DM degradability of corn silage and CP degradability of concentrate increased with RPP supplementation and tended to increase with increasing dietary CP levels, while NDF degradability of corn silage and DM degradability of concentrate increased with increasing dietary CP levels or RPP supplementation. Activities of carboxymethyl-cellulase, cellobiase, xylanase, pectinase, α-amylase and protease, populations of Ruminococcus albus, Ruminococcus flavefaciens, Butyrivibrio fibrisolvens and Prevotella ruminicola, and urinary excretion of purine derivatives increased with increasing dietary CP levels or RPP supplementation, but populations of Fibrobacter succinogenes and R. amylophilus increased with RPP supplementation, and decreased with increasing dietary CP levels. The results showed that increasing dietary CP levels from 113.7g to 133.9g or with 0.48g RPP supplementation improved ruminal fermentation and microbial protein synthesis by increasing microbial populations and enzymes activities.

Effect of ginkgo extract supplementation on in vitro rumen fermentation and bacterial profiles under different dietary conditions.

Ginkgo extract was applied to a batch culture study and evaluated for its potential as a feed additive for ruminant animals under different forage-to-concentrate (F:C) ratios (1:9, 3:7, 5:5, 7:3 and 9:1). Rumen fluid was mixed with respective diet and incubated at 39°C for 24h with and without ginkgo extract (1.6% fruit equivalent in culture). Methane production was significantly decreased by ginkgo extract, with the greatest reductions found in the 5:5 (41.9%) followed by the 7:3 ratios (36.7%). Total short chain fatty acid and ammonia levels were not affected by ginkgo extract supplementation in any of the five different diets. However, ginkgo extract increased propionate proportion and decreased acetate proportion in all dietary conditions tested. The levels of total bacteria, Ruminococcus flavefaciens, Ruminococcus albus and Fibrobacter succinogenes were decreased by ginkgo extract. The levels of Selenomonas ruminantium, Anaerovibrio lipolytica, Ruminobacter amylophilus, Succinivibrio dextrinosolvens and Megasphaera elsdenii were increased by ginkgo extract supplementation, possibly contributing to the higher propionate production. These results suggest that rumen modulation by ginkgo extract can be achieved at a wide range of F:C ratios with no adverse impact on feed digestion. Moreover, F:C ratios of 5:5 and 7:3 may be optimal when methane mitigation is expected.