Milk odd- and branched-chain fatty acids in relation to the rumen fermentation pattern

The objectives of this study were 1) to determine whether a relationship exists between molar proportions of volatile fatty acids in the rumen and milk odd-and branched-chain fatty acid concentrations (i.e., iso C13:0, anteiso C13:0, iso C14:0, C15:0, iso C15:0, anteiso C15:0, iso C16:0, C17:0, iso C17:0, anteiso C17:0, and cis-9 C17:1); and 2) to evaluate the accuracy of prediction of the latter equations using an independent data set. For development of the regression equations, individual cow data from 10 feeding experiments with rumen-fistulated dairy cows were used, resulting in a data set of 148 observations. Milk odd- and branched-chain fatty acids were closely related to the molar proportions of acetate (SE = 15.3 mmol/mol), propionate (SE = 14.7 mmol/mol), and butyrate (SE = 9.2 mmol/mol). These regression equations were further validated using data from the literature (n = 14). Evaluation of these prediction equations using the independent data set resulted in a root mean square prediction error of 3.0, 9.0, and 8.9% of the observed mean for acetate, propionate, and butyrate, respectively. In addition, less then 5% of the mean square prediction error was due to line bias. This suggests that the currently developed prediction equations based on milk odd- and branched-chain fatty acids show potential to predict molar proportions of individual volatile fatty acids in the rumen.     

Effects of extracts of spices on rumen methanogenesis,enzyme activities and fermentation of feeds in vitro

BACKGROUND: An experiment was conducted to study the effects of boiling water, methanol and ethanol extracts (0, 0.25 and 0.50 mL) of seeds of Foeniculum vulgare (fennel), flower buds of Syzygium aromaticum (clove), bulbs of Allium sativum (garlic), bulbs of Allium cepa (onion) and roots of Zingiber officinalis (ginger) on rumen methanogenesis, fibrolytic enzyme activities and fermentation characteristics in vitro. RESULTS: Ethanol and methanol extracts of fennel, clove and garlic at 0.50 mL and clove at 0.25 mL inhibited (P < 0.05) methane production. Carboxymethylcellulase activity was reduced (P < 0.05) by ethanol and methanol extracts (0.50 mL) of fennel and clove (0.25 and 0.50 mL). The extracts of clove reduced (0.25 and 0.50 mL) xylanase and acetylesterase activities, and the fennel extract (0.50 mL) reduced (P < 0.05) xylanase activity. However, the extracts of garlic (0.50 mL) increased (P < 0.05) acetylesterase activity. Concentrations of volatile fatty acids were reduced (P < 0.05) by the extracts of garlic and onion. The extracts of garlic caused a decrease (P < 0.05) in acetate:propionate ratio (A:P) at 0.50 mL, whereas A:P was increased (P < 0.05) by the inclusion of 0.50 mL extracts of clove. Methanol and ethanol extracts of clove decreased (P < 0.05) in vitro organic matter degradability. Extracts (0.50 mL) of clove decreased (P < 0.05) the numbers of total protozoa, small entodiniomorphs and holotrichs, whereas extracts of onion, ginger and garlic enhanced (P < 0.05) protozoal numbers (both entodiniomorphs and holotrichs). CONCLUSION: Ethanol and methanol extracts of fennel and garlic have potential to inhibit rumen methanogenesis without adversely affecting rumen fermentation. Copyright © 2009 Society of Chemical Industry     

Proper motility enhances rumen fermentation and microbial protein synthesis with decreased saturation of dissolved gases in rumen simulation technique

The physiological function of the reticulorumen plays an essential role in ruminant nutrition, and detailed knowledge of rumen motility can further advance understanding of ruminant nutrition and physiology. Rumen motility was simulated by setting different stirrer rotation speeds in a rumen simulation technique (RUSITEC) system. The aim of this study was to investigate the effects of rotation speeds on rumen fermentation, saturation factor of dissolved gases, hydrogen (H₂) and methane (CH₄) emissions, microbial protein synthesis, and selected microbial population using RUSITEC. The experiment was performed according to a balanced 3 × 3 Latin square design, and each period included 7 d for adaptation and 3 d for sampling. Three motility treatments included 5, 15, and 25 rpm rotation speeds. Daily total gas and H₂ and CH₄ emissions had quadratic responses to the increasing rotation speed and were highest at 15 rpm. Quadratic and linear responses (highest at 5 rpm) to increasing rotation speed were observed for saturation factors of H₂ and CH₄, liquid-dissolved H₂ and CH₄ concentrations, and headspace concentration of H₂ in the gas phase, whereas increasing rotation speed linearly decreased saturation factors of CO₂ and liquid-dissolved CO₂ concentration. Quadratic and linear responses to increasing rotation speed were observed for molar percentages of acetate, ammonia, and microbial protein concentration, whereas increasing rotation speed quadratically increased pH and decreased total volatile fatty acid concentration and acetate-to-propionate ratio. The 15-rpm rotation speed had the highest values of total volatile fatty acids, acetate molar percentage, and microbial protein concentration. Quadratic and linear responses to increasing rotation speed were observed for copy numbers of solid-associated fungi and fluid-associated bacteria, fungi, and protozoa, while increasing rotation speed linearly increased copy numbers of solid-associated protozoa. Rotation at 15 rpm increased populations of fungi and protozoa in the solid rumen contents and the population of bacteria and fungi in the liquid rumen contents. In summary, this study provides insights on the biofunction of proper rumen motility (i.e., at a rotation speed of 15 rpm), such as improving feed fermentation, increasing gas emissions with decreased dissolved gas concentrations and saturation factors, and promoting microbial colonization and microbial protein synthesis, although further increase in rotation speed (i.e., to 25 rpm) decreases feed fermentation and microbial protein synthesis.     

Effects of Cordyceps militaris on the growth of rumen microorganisms and in vitro rumen fermentation with respect to methane emissions

This experiment was designed to investigate the effects of different concentrations (0.00, 0.10, 0.15, 0.20, 0.25, and 0.30 g/L) of dried Cordyceps militaris mushroom on in vitro anaerobic ruminal microbe fermentation and methane production using soluble starch as a substrate. Ruminal fluids were collected from Korean native cattle, mixed with phosphate buffer (1:2), and incubated anaerobically at 38°C for 3, 6, 9, 12, 24, 36, 48, and 72 h. The addition of C. militaris significantly increased total volatile fatty acid and total gas production. The molar proportion of acetate was decreased and that of propionate was increased, with a corresponding decrease in the acetate:propionate ratio. As the concentration of C. militaris increased from 0.10 to 0.30 g/L, methane and hydrogen production decreased. The decrease in methane accumulation relative to the control was 14.1, 22.0, 24.9, 39.7, and 40.9% for the 0.10, 0.15, 0.20, 0.25, and 0.30 g/L treatments, respectively. Ammonia-N concentration and numbers of live protozoa decreased linearly with increasing concentrations of C. militaris. The pH of the medium significantly decreased at the highest level of C. militaris compared with the control. In conclusion, C. militaris stimulated mixed ruminal microorganism fermentation and inhibited methane production in vitro. Therefore, C. militaris could be developed as a novel compound for antimethanogenesis.     

Plant extracts affect in vitro rumen microbial fermentation

Different doses of 12 plant extracts and 6 secondary plant metabolites were incubated for 24 h in diluted ruminal fluid with a 50:50 forage:concentrate diet. Treatments were: control (no additive), plant extracts (anise oil, cade oil, capsicum oil, cinnamon oil, clove bud oil, dill oil, fenugreek, garlic oil, ginger oil, oregano oil, tea tree oil, and yucca), and secondary plant metabolites (anethol, benzyl salicylate, carvacrol, carvone, cinnamaldehyde, and eugenol). Each treatment was supplied at 3, 30, 300, and 3,000 mg/L of culture fluid. At 3,000 mg/L, most treatments decreased total volatile fatty acid concentration, but cade oil, capsicum oil, dill oil, fenugreek, ginger oil, and yucca had no effect. Different doses of anethol, anise oil, carvone, and tea tree oil decreased the proportion of acetate and propionate, which suggests that these compounds may not be nutritionally beneficial to dairy cattle. Garlic oil (300 and 3,000 mg/L) and benzyl salicylate (300 and 3,000 mg/L) reduced acetate and increased propionate and butyrate proportions, suggesting that methane production was inhibited. At 3,000 mg/L, capsicum oil, carvacrol, carvone, cinnamaldehyde, cinnamon oil, clove bud oil, eugenol, fenugreek, and oregano oil resulted in a 30 to 50% reduction in ammonia N concentration. Careful selection and combination of these extracts may allow the manipulation of rumen microbial fermentation.     

Effect of forage:concentrate ratio on fatty acid composition of rumen bacteria isolated from ruminal and duodenal digesta

Four dairy cows were used to examine the effect of the dietary forage:concentrate ratio [35:65, 50:50, 65:35, and 80:20 on a dry matter (DM) basis] on the fatty acid composition of rumen bacteria isolated from the liquid (LAB) and solid (SAB) phase of the rumen and duodenal digesta. Rumen contents were sampled 4 h after the morning feeding. Solid and liquid phases were separated from rumen contents and duodenal bacteria from a composite duodenal sample by differential centrifugation. Total fatty acid content in bacterial DM was 1.6 to 2.8 times higher in SAB compared with LAB, and increased with dietary concentrate. In combination with published reports, the data show that bacterial fatty acid content and composition is closely related to dietary fatty acids except for C18:2n-6 and C18:3n-3. A decrease in forage:concentrate ratio increased bacterial concentration of trans-10 C18:1, and this increase was 3.4 times higher in LAB compared with SAB. Analysis of odd- and branched-chain fatty acids showed large differences between SAB and LAB, which probably reflected a difference in species composition. The variation in odd- and branched-chain fatty acids between SAB and LAB was used to estimate their relative proportions in duodenal bacteria by means of linear programming, and showed an increased proportion of SAB from 64.7 to 74.8% with increasing forage:concentrate ratio. In addition, increasing the proportion of dietary forage was closely related to the proportion of anteiso C15:0 in total odd- and branched-chain fatty acids (r_pearson = −0.771). The bacterial concentration of iso C17:0 closely reflected the bacterial growth rate as shown by the relation with cytosine:N (r_pearson = −0.729). These strong relationships suggest that odd- and branched-chain fatty acids might be used as tool to evaluate nutrient supply to rumen bacteria.     

Effect of essential oil active compounds on rumen microbial fermentation and nutrient flow in in vitro systems

Two experiments were conducted to determine the effects of several essential oil active compounds on rumen microbial fermentation. In the first experiment, 4 doses (5, 50, 500, and 5,000 mg/L) of 5 essential oil compounds were evaluated using in vitro 24-h batch culture of rumen fluid with a 60:40 forage:concentrate diet (18% crude protein; 30% neutral detergent fiber). Treatments were control (CON), eugenol (EUG), guaiacol, limonene, thymol (THY), and vanillin. After 24 h, the pH was determined, and samples were collected to analyze ammonia N and volatile fatty acids (VFA). The highest dose of all compounds decreased total VFA concentration and increased the final pH. Eugenol at 5 mg/L tended to reduce the proportion of acetate and the acetate to propionate ratio, at 50 and 500 mg/L tended to reduce ammonia N concentration, and at 500 mg/L reduced the proportion of propionate and branched-chain VFA concentration, without affecting total VFA concentration. All other treatments had minor effects or changes occurred only after total VFA concentration decreased. In the second experiment, 8 dual-flow continuous culture fermenters (1,320 mL) were used in 3 replicated periods (6 d of adaptation and 3 d of sampling) to study the effects of THY and EUG on rumen microbial fermentation. Fermenters were fed 95 g/d of DM of a 60:40 forage:concentrate diet (18% crude protein; 30% neutral detergent fiber). Treatments were CON, 10 mg/L of monensin (positive control), and 5, 50, or 500 mg/L of THY and EUG, and were randomly assigned to fermenters within periods. During the last 3 d of each period, samples were taken at 0, 2, 4, and 6 h after the morning feeding and analyzed for peptides, amino acids, and ammonia N concentrations, and total and individual VFA concentrations. Monensin changed the VFA profile as expected, but inhibited nutrient digestion. Eugenol and THY decreased total VFA concentration and changed the VFA profile, and only 5 mg/L of THY tended to reduce the proportion of acetate, increased the proportion of butyrate, and increased the large peptides N concentration without decreasing total VFA concentration. Most of these essential oil compounds demonstrated their antimicrobial activity by decreasing total VFA concentration at high doses. However, EUG in batch fermentation and 5 mg/L of THY in continuous culture modified the VFA profile without decreasing total VFA concentration, and EUG in batch fermentation decreased ammonia N concentration.     

Effect of a monensin controlled-release capsule on rumen and blood metabolites in Florida Holstein transition cows

The objective of this study was to determine the effect of a monensin controlled-release capsule administered intraruminally at dry-off on rumen volatile fatty acids, NH3, lactate, pH, and energy blood metabolites in transition dairy cows fed Florida typical diets. In March 2003, 24 cows (10 primiparous and 14 multiparous) dried-off 50 to 70 d before expected parturition were randomly assigned to a treatment (n=12, oral capsule of monensin) or a control group (n=12, no capsule). Both groups received the same diet and were exposed to the same environment and management conditions. At assignment, at 21 d before expected parturition, at calving, and at 7, 14, and 21 d postpartum, blood samples were taken and body condition scores were determined. At 10 d postpartum, rumen and blood samples were obtained in the morning before the first feeding and at 2, 4, and 6 h after feeding. Serum nonesterified fatty acids (NEFA), beta-hydroxybutyrate (BHBA), and glucose were measured. Rumen samples were analyzed for concentrations of acetic, propionic, butyric, L- and D-lactic acids, and NH3. Data for rumen and blood metabolites were analyzed by ANOVA, mixed models for repeated measures. Volatile fatty acids were not different between groups. Multiparous treated cows had a significant reduction in rumen NH3 at 6 h after feeding. Treatment with monensin significantly increased body condition score at calving in multiparous cows. During the postpartum period, NEFA and BHBA were noticeably lower in treated than in control primiparous cows. This difference was not observed in multiparous cows.     

Amino acid composition of rumen bacteria and protozoa in cattle

Because microbial crude protein (MCP) constitutes more than 50% of the protein digested in cattle, its AA composition is needed to adequately estimate AA supply. Our objective was to update the AA contributions of the rumen microbial AA flowing to the duodenum using only studies from cattle, differentiating between fluid-associated bacteria (FAB), particle-associated bacteria (PAB), and protozoa, based on published literature (53, 16, and 18 treatment means were used for each type of microorganism, respectively). In addition, Cys and Met reported concentrations were retained only when an adequate protection of the sulfur groups was performed before the acid hydrolysis. The total AA (or true protein) fraction represented 82.4% of CP in bacteria. For 10 AA, including 4 essential AA, the AA composition differed between protozoa and bacteria. The most noticeable differences were a 45% lower Lys concentration and 40% higher Ala concentration in bacteria than in protozoa. Differences between FAB and PAB were less pronounced than differences between bacteria and protozoa. Assuming 33% FAB, 50% PAB, and 17% of protozoa in MCP duodenal flow, the updated concentrations of AA would decrease supply estimates of Met, Thr, and Val originating from MCP and increase those of Lys and Phe by 5 to 10% compared with those calculated using the FAB composition reported previously. Therefore, inclusion of the contribution of PAB and protozoa to the duodenal MCP flow is needed to adequately estimate AA supply from microbial origin when a factorial method is used to estimate duodenal AA flow. Furthermore, acknowledging the fact that hydrolysis of 1 kg of true microbial protein yields 1.16 kg of free AA substantially increases the estimates of AA supply from MCP.     

Effect of inoculated corn silage enriched with sunflower oil on rumen fermentation and lipid metabolism in an artificial rumen (RUSITEC)

BACKGROUND: Some rumen isolates are able to produce conjugated linoleic acid (CLA) from linoleic acid (LA) in vitro. Effects of providing diets containing corn silage (CS) and lucerne hay to an artificial rumen (RUSITEC) in which the corn was not inoculated (CS), or inoculated with Lactobacillus plantarum CCM 4000 (CS + LP), Lactobacillus fermentum LF2 (CS + LF) or Enterococcus faecium CCM 4231 (CS + EF) and supplied with sunflower oil (SO; 30 g kg^?1; w/w) on rumen metabolism were examined. RESULTS: The SO affected the outputs of all fatty acids. TVA output of uninoculated CS with SO was lower as compared to inoculated CS. The interaction of the CS × SO in the daily output of TVA was detected (P < 0.001). The biohydrogenation of oleic, linoleic, α‐linolenic and total fatty acids was influenced by SO (P < 0.05 and P < 0.001). CONCLUSION: Inoculated silage induces changes in the rumen metabolism which might be related to differences observed in the extent of rumen BH of PUFA in RUSITEC. SO supplementation might positively enhances the production of some rumen intermediates; however, relationships between inoculated silages and oil supplementation can be presumed in the daily production of trans‐vaccenic acid. Copyright © 2009 Society of Chemical Industry     

Disappearance of Maillard reaction products during ensilage and rumen fermentation in vitro

The disappearance of Maillard reaction products (MRPs) during ensilage and rumen fermentation was studied. MRPs were prepared by heating mixtures of D ‐glucose and glycine (GG) or D ‐xylose and glycine (XG); lyophilised powder (MW > 1000) was subjected to anaerobic incubation for 24 h with lucerne juice or buffered rumen fluid. Changes in MRPs were assessed by comparing gel filtration profiles before and after incubation, and the proportion of disappeared MRPs was calculated based on the area under the curve. A portion of MRPs, particularly high‐MW fractions, disappeared when incubated with lucerne juice, and the extent was greater (P < 0.01) in XG than in GG. The disappearance of GG was increased (P < 0.01) when ensilage was fortified with added glucose, while that of XG was unaffected with or without the fermentable substrate. Rumen fermentation had little impact on GG and XG, whereas a portion of MRPs disappeared when microbial activity was enhanced by adding lucerne hay to the media. Volatile fatty acid production was unaffected by MRPs in either ensilage or rumen fermentation. These results suggest that the effect on MRPs of anaerobic fermentation may be different between ensilage and rumen digestion. Significant amounts of MRPs would enter the intestine when ruminants are fed hay‐based diets rather than silage‐based diets. © 2000 Society of Chemical Industry     

Rates of production of acetate, propionate, and butyrate in the rumen of lactating dairy cows given normal and low-roughage diets

Five lactating dairy cows with a permanent cannula in the rumen were given (kg DM/d) a normal diet (7.8 concentrates, 5.1 hay) or a low-roughage (LR) diet (11.5 concentrates, 1.2 hay) in two meals daily in a two-period crossover design. Milk fat (g/kg) was severely reduced on diet LR. To measure rates of production of individual volatile fatty acids (VFA) in the rumen, 0.5 mCi 1-(14)C-acetic acid, 2-(14)C-propionic acid, or 1-(14)C-n-butyric acid were infused into the rumen for 22 h at intervals of 2 to 6 d; rumen samples were taken over the last 12 h. To measure rumen volume, we infused Cr-EDTA into the rumen continuously, and polyethylene glycol was injected 2 h before the morning feed. Results were very variable, so volumes measured by rumen emptying were used instead. Net production of propionic acid more than doubled on LR, but acetate and butyrate production was only numerically lower. Net production rates pooled across both diets were significantly related to concentrations for each VFA. Molar proportions of net production were only slightly higher than molar proportions of concentrations for acetate and propionate but were lower for butyrate. The net energy value (MJ/d) of production of the three VFA increased from 89.5 on normal to 109.1 on LR, equivalent to 55 and 64% of digestible energy, respectively. Fully interchanging, three-pool models of VFA C fluxes are presented. It is concluded that net production rates of VFA can be measured in non-steady states without the need to measure rumen volumes.     

Effect of chemical form,heating, and oxidation products of linoleic acid on rumen bacterial population and activities of biohydrogenating enzymes

Heating polyunsaturated fatty acids (PUFA) produces oxidation products, such as hydroperoxides, aldehydes, and oxypolymers, which could be responsible at least in part for modification of PUFA rumen biohydrogenation (BH). Three in vitro experiments were conducted to investigate the effects of linoleic acid (cis-9,cis-12-C18:2) oxidation products on BH. In the first experiment, we studied the effects of free linoleic acid (FLA), heated FLA (HFLA, at 150°C for 6 h), triacylglycerols of linoleic acid (TGLA), heated TGLA (HTGLA, at 150°C for 6 h), 13-hydroperoxide (13HPOD), trans-2-decenal (T2D), and hexanal (HEX) on BH in vitro after 6 and 24 h of incubation. In the second experiment, aldehydes differing in chain length and degree of unsaturation [pentanal, HEX, heptanal, nonanal, T2D, trans-2,trans-4-decadienal (T2T4D)] were incubated in vitro for 5 h in rumen fluid. In the third experiment, 9-hydroperoxide (9HPOD), 13HPOD, HEX, or T2T4D were incubated for 1 h in rumen fluid inactivated with chloramphenicol to investigate their effects on enzyme activity. In experiment 1, heat treatment of TGLA generated TGLA oxypolymers, did not affect cis-9,cis-12-C18:2 disappearance, but did decrease BH intermediates, especially trans-11 isomers. Heating FLA decreased cis-9,cis-12-C18:2 disappearance and cis-9,trans-11-CLA and trans-11-C18:1 production. Treatment with HEX and T2D did not affect cis-9,cis-12-C18:2 disappearance and barely affected production of BH intermediates. The bacterial community was affected by 13HPOD compared with FLA and HFLA, in parallel with an increase in trans-10 isomer production after a 6-h incubation. After 24 h of incubation, 13HPOD decreased trans-11 isomer production, but to a lesser extent than HFLA. In experiment 2, some weak but significant effects were observed on BH, unrelated to chain length or degree of unsaturation of aldehydes; the bacterial community was not affected. In experiment 3, 9HPOD inhibited Δ⁹-isomerization, and both 9HPOD and 13HPOD inhibited Δ¹²-isomerization. We concluded that oxypolymers did not affect cis-9,cis-12-C18:2 disappearance. Heating both esterified and free cis-9,cis-12-C18:2 greatly altered Δ¹²-isomerization. Aldehydes had few effects. Hydroperoxides are responsible, at least in part, for the effects of fat heating: 13HPOD increased trans-10 isomer production (probably by affecting the bacterial community) and decreased trans-11 isomer production by inhibiting Δ¹²-isomerase activity, whereas 9HPOD inhibited both isomerases.     

Effect of calcium salts of fatty acids on rumen function and the digestibility of rations by sheep

Experiments were conducted to determine the effects of supplementing chopped ryegrass hay with 0, 1.5, 3, 6 or 12% calcium salts of fatty acids (CaSFA; Megalac) on digestion in the rumen of cannulated cows and on nutrient digestibility by sheep. In-situ ruminal disappearances of dry matter (DM) and cell wall fractions (acid detergent fibre, ADF, and neutral detergent fibre, NDF) were measured by the nylon bag technique. The DM, ADF and NDF disappearances were reduced for diets containing CaSFA at the 1.5, 3 or 6% levels after 48 h of rumen exposure. For diets containing 12% CaSFA, irrespective of the time of rumen incubation, DM disappearance was decreased, while ADF and NDF breakdown was improved. CaSFA supplementation did not influence invitro fermentation characteristics (pH, volatile fatty acid patterns). A digestibility experiment with six mature sheep showed no difference in nitrogen digestibility between unsupplemented and CaSFA-supplemented diets. However, digestibilities of DM, ADF and NDF were higher relative to the basal for the 12% CaSFA: 63.1 versus 60.6%, 63.8 versus 58.7% and 67.4 versus 62.7%, respectively. Ether extract digestion was enhanced by increasing the level of CaSFA, the corresponding values being 49.1, 66.5, 78.6, 81.9 and 77.9% at 0, 1.5, 3, 6 and 12% CaSFA, respectively. Apparent digestibility of energy was higher for the 12% CaSFA than the control diet 68.8 versus 62.1%). Nitrogen and energy retention was improved due to the decreases in urine and faecal excretion, respectively.     

Changes in rumen fermentation and bacterial community in lactating dairy cows with subacute rumen acidosis following rumen content transplantation

Rumen microbiota intervention has long been used to cure ruminal indigestion in production and has recently become a research hotspot. However, how it controls the remodeling of rumen bacterial homeostasis and the restoration of rumen fermentation in cows of subacute ruminal acidosis (SARA) remains poorly understood. This study explored changes in rumen fermentation and bacterial communities in SARA cows following rumen content transplantation (RCT). The entire experiment comprised 2 periods: the SARA induction period and the RCT period. During the SARA induction period, 12 ruminally cannulated lactating Holstein cows were selected and allocated into 2 groups at random, fed either a conventional diet [CON; n = 4; 40% concentrate, dry matter (DM) basis] or a high-grain diet (HG; n = 8; 60% concentrate, DM basis). After the SARA induction period, the RCT period began. The HG cows were randomly divided into 2 groups: the donor-recipient (DR) group and the self-recipient (SR) group, and their rumen contents were removed completely before RCT. For the DR group, cows received 70% rumen content from the CON cows, paired based on comparable body weight; for the SR group, each cow received 70% rumen content, self-derived. The results showed that nearly all rumen fermentation parameters returned to the normal levels that the cows had before SARA induction after 6 d of transplantation, regardless of RCT. The concentrations of acetate, valerate, and total volatile fatty acids (VFA) were not recovered in the SR cows, whereas all of them were recovered in the DR cows. The amplicon sequencing results indicated that both the SR and DR cows rebuild their rumen bacterial homeostasis quickly within 4 d after RCT, and the DR group showed a higher level of bacterial community diversity. At the genus level, the DR cows displayed an improved proportion of unclassified Ruminococcaceae and Saccharofermentans compared with the SR cows. Correlation analysis between the rumen bacteria and rumen fermentation suggested some potential relationships between the predominant transplantation-sensitive operational taxonomic units and VFA. Co-occurrence network analysis revealed that RCT affected only those rumen bacterial taxa that showed weak interactions with other taxa and did not affect the pivotal rumen bacteria with high levels of co-occurrence. Our findings indicate that RCT contributes to the restoration of rumen bacterial homeostasis and rumen fermentation in cows suffering from SARA without affecting the core microbiome.     

Effects of citric acid supplementation on rumen fermentation,urinary excretion of purine derivatives and feed digestibility in steers

BACKGROUND: An increase feed efficiency and rate of gain were observed from addition of sodium citrate to the diet of fatting lamb. It is suggested that the small amounts of citric acid (CA) may have a catalytic effect on rumen microbial metabolism which would result in a potential means of increasing feed efficiency. The objective was to evaluate the effects of CA supplementation on rumen fermentation, ruminal microbial production by measuring urinary excretion of purine derivatives, and digestibility in the total tract of steers. RESULTS: Ruminal pH linearly (P = 0.01) decreased, whereas total volatile fatty acid concentration linearly (P = 0.01) increased with increasing CA supplementation. Ratio of acetate to propionate linearly (P = 0.01) increased due to the increase in acetate production. Urinary excretion of purine derivatives was quadratically (P = 0.02) changed, with the lowest for control, medium for low CA and highest for medium and high CA supplementation. Similarly, digestibilities of nutrients in the total tract were also linearly and quadratically increased with increasing dosages of CA. CONCLUSION: Supplementation of CA increased rumen acetate concentration and thus increased ratio of acetate to propionate. Urinary excretion of purine derivatives and total digestibility were improved. In the experimental conditions of this trial, the optimum citric acid dose was about 200 g citric acid per steer per day. Copyright © 2009 Society of Chemical Industry     

Short communication: Sodium salicylate negatively affects rumen fermentation in vitro and in situ

Administration of sodium salicylate (SS) to cows in early lactation has a positive effect on whole-lactation milk production but a negative effect on metabolism in some cases. The objective of this trial was to determine whether SS directly affects rumen fermentation. Experiment 1 was designed to investigate the effects of direct inclusion of SS in a 24-h batch culture, and experiment 2 was designed to test the fermentative ability of rumen fluid from heifers who had received SS. In experiment 1, we combined strained and pooled rumen fluid from 3 heifers in a 2:1 ratio with McDougall's buffer, and added 150 mL of the inoculum to each flask (n = 5/treatment) with 2.5 g of fermentation substrate similar to a lactating cow ration, ground to 1 mm. We then added premixed treatments (1-mL volume) to achieve the desired final amount of SS (CON1 = 0 mg, LOW = 125 mg, MED = 250 mg, HI = 375 mg). In experiment 2, 6 heifers (n = 3/treatment) were drenched daily for 3 d, either with 62.5 g of SS dissolved in water (SAL) or an equal volume of water (CON2). Rumen fluid was collected from each heifer and was not pooled. After the fluid was mixed 2:1 with McDougall's buffer, 150 mL of inoculum was added to the fermentation flasks (n = 4/heifer) with 2.5 g of fermentation substrate. This experiment was performed the day before SS treatment began and repeated 1, 13, and 35 d after the end of the treatment period. We also performed an in situ experiment at each of these time points. In the first experiment, inclusion of SS resulted in a decrease in dry matter disappearance (DMD) over 24 h, as well as an increase in final pH. We detected no difference between treatments for gas production asymptotic volume, rate, or lag. In the second experiment, we detected a significant treatment × day interaction for DMD: we observed no difference between groups during a 24-h batch culture on the day following treatment, but SAL resulted in lower DMD on d 13 and d 35. We detected no treatment effect on the final pH of the batch culture or on any gas-production parameters. We observed a tendency for SAL to decrease the DMD rate in situ on the day after treatment. These results indicate that SS administration has a negative effect on rumen microorganisms.     

Rumen volatile fatty acid molar proportions,rumen epithelial gene expression,and blood metabolite concentration responses to ruminally degradable starch and fiber supplies

The objective of this work was to characterize rumen volatile fatty acid (VFA) concentrations, rumen epithelial gene expression, and blood metabolite responses to diets with different starch and fiber sources. Six ruminally cannulated yearling Holstein heifers (body weight = 330 ± 11.3 kg) were arranged in a partially replicated Latin square experiment with 4 treatments consisting of different starch [barley (BAR) or corn (CRN)] and fiber [timothy hay (TH) or beet pulp (BP)] sources. Treatments were arranged as a 2 × 2 factorial. Beet pulp and TH were used to create relative changes in apparent ruminal fiber disappearance, whereas CRN and BAR were used to create relative changes in apparent ruminal starch disappearance. Each period consisted of 3 d of diet adaptation and 15 d of dietary treatment. In situ disappearance of fiber and starch were estimated from bags incubated in the rumen from d 10 to 14. From d 15 to 17, rumen fluid was collected every hour from 0500 to 2300 h. Rumen fluid samples were pooled by animal/period and analyzed for pH and VFA concentrations. On d 18, 60 to 80 papillae were biopsied from the epithelium and preserved for gene expression analysis. On d 18, one blood sample per heifer was collected from the coccygeal vessel. In situ ruminal starch disappearance rate (7.30 to 8.72%/h for BAR vs. 7.61 to 10.5%/h for CRN) and the extent of fiber disappearance (22.2 to 33.4% of DM for TH vs. 34.4 to 38.7% of DM for BP) were affected by starch and fiber source, respectively. Analysis of VFA molar proportions showed a shift from propionate to acetate, and valerate to isovalerate on TH diets compared with BP. Corn diets favored propionate over butyrate in comparison to BAR diets. Corn diets also had higher molar proportions of valerate. Expression of 1 gene (SLC9A3) were increased in BP diets and 2 genes (BDH1 and SLC16A4) tended to be increased in TH diets. Plasma acetate demonstrated a tendency for a starch by fiber interaction with BAR-BP diets having the highest plasma acetate, but other metabolites measured were not significant. These results suggest that TH has the greatest effect on shifts in VFA molar proportions and epithelial transporters, but does not demonstrate shifts in blood metabolite concentrations.