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Dietary tannins can affect rumen microbiota and enteric fermentation to mitigate methane emissions, although such effects have not yet been fully elucidated. We tested two subunits of hydrolyzable tannins named gallic acid (GA) and ellagic acid (EA), alone (75 mg/g DM each) or combined (150 mg/g DM in total), using the Rusitec system. EA and EA+GA treatments decreased methane production, volatile fatty acids, nutrient degradation, relative abundance of Butyrivibrio fibrisolvens , Fibrobacter succinogenes , Ruminococcus flavefaciens but increased Selenomonas ruminantium . EA and EA+GA increased urolithins A and B. Also, EA and EA+GA reduced bacterial richness, with limited effects on archaeal richness. For bacteria, Megasphaera elsdenii was more abundant after EA and EA+GA, while Methanomethylophilaceae dominated archaea in all treatments. EA was more effective than GA in altering rumen microbiota and fermentation but GA did not reduce VFA and nutrient degradation. Thus, dietary supplementation of EA-plant extracts for ruminants may be considered to mitigate enteric methane, although a suitable dosage must be ensured to minimize the negative effects on fermentation.

Barley grain sources with variable kernel sizes makes adequate and consistent processing of kernels challenging. This study evaluated how the severity of processing for reconstituted high-moisture (65% on DM basis) barley (RHB) affects ensiling characteristics and in vitro ruminal fermentation. Three independent sources of light (<630 g/500 mL) and heavy (>670 g/500 mL) barley were blended to create 4 sources of variable kernel sized barley (646 g/500 mL). Reconstituted high-moisture barley rolled using a 1.40-mm gap width for finely processed RHB (RHBF), a 1.86-mm gap width for medium processed RHB (RHBM), or a 2.31-mm gap width for coarsely processed RHB (RHBC) was ensiled for 1 or 5 mo with dry rolled barley (DRB; roller gap width 1.86 mm) used as a control. The 1-mo RHB and the DRB were further evaluated using the rumen in vitro simulation technique (RUSITEC) to investigate the effects of severity of processing for RHB on ruminal fermentation, and gas, methane, and microbial protein production. Using a randomized complete block design (n = 4), 16 fermenters from 2 RUSITEC apparatuses were used to assess the 4 sources and 4 processing treatments. The addition of water increased kernel width before rolling and resulted in increased kernel length, width, and thickness for RHB relative to DRB. Increasing processing severity for RHB linearly increased kernel width. The percentage of fine particles (<1.18 mm) was greater for DRB than RHBF, but did not differ by processing severity for RHB. Dry matter, OM, and starch disappearance were not different between DRB and RHBF, but linearly increased with increasing processing severity for RHB. Fermenter pH tended to be less for DRB relative to RHBF. In conclusion, the reduction in fine particles with the addition of water for RHB may prevent a decline in fermenter pH and when processed to achieve the same processing index using a smaller roller gap width, yielded similar DM and OM disappearance suggesting a lesser risk for low ruminal pH without compromising digestibility., (© 2024, The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Objectives: Feeding winery by-products (WBP) could affect the bovine microbiome because of their phenol compounds and a transfer of WBP-associated microbiota. This work examined changes in the underexplored solid-associated rumen microbiome following the inclusion of WBP., Methods: Using the rumen simulation technique, fermenters were inoculated with the inoculum of donor cows and were fed one of six dietary treatments including a control diet of 70 % hay +30 % concentrate (CON), control diet + 3.7 % commercial grapeseed extract (EXT), 65 % hay + 25 % concentrate + 10 % grape pomace (GP-low), 56 % hay + 24 % concentrate + 20 % grape pomace (GP-high), 70 % hay + 25 % concentrate + 5 % grapeseed meal (GS-low), and 65 % hay + 25 % concentrate + 10 % grapeseed meal (GS-high) (dry matter basis). The compositional changes of bacteria, archaea and fungi in the solid fractions were based on 16S and ITS2 rRNA sequencing., Results: The alpha- and beta-diversity of the microbiota were unaffected. However, treatment modified the bacterial composition at low taxonomic levels. Butyrivibrio fibrisolvens, Treponema bryantii, and bacterium MC2010 decreased in EXT, while Treponema berlinense was increased in GP-high and GP-low compared to CON. Concerning fungi, GS-high increased Candida spp., Lachancea spp., Microdochium spp., Mucor spp., Pichia spp., Saturnispora spp., and Zygosaccharomyces spp. compared to CON. Many non-Saccharomyces yeasts were detected in WBP samples but absent in donor cows and CON samples. The genera affected by treatment were not the major contributors to the ruminal degradation of nutrients., Conclusions: The results indicate a sensitivity of rumen solid bacteria to grape phenols when delivered as an extract and a transfer of WBP-associated microbiota into the rumen., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The approving agencies for plant protection agents request xenobiotic metabolism and residue studies in rats, farm animals and plants (e.g. EU regulation 1107/2009) according to OECD guidelines. The specific intestinal physiology of ruminants might lead to specific residues, which should be investigated very carefully. Specific aspects of xenobiotic metabolism in ruminants may arise, which are investigated by performing additional in vivo studies. The aim of the present work is to asses a modified rumen simulation system (RUSITEC) for such studies in vitro. Rumen constituents from sheep were incubated over 8 days. Physiological parameters followed by monitoring the pH (6.70 ± 0.07), the redox potential (301 mV ± 30 mV), the microbial composition and determination of β-glucosidase activity. After anequilibration period of three to four days the fermenters were probed with 14C-labelled triazolederivatives, i.e.common metabolites of azole fungicides. Only triazole-alanine was cleaved to 1,2,4-triazole, while triazole-acetic acid and triazole-lactic acid remained stable up to 96 h. Moreover, glycosides are often determined as the main residues in the plants. This analysis showed, that the two glucosides octyl-β-D-glucopyranoside and polydatin were both rapidly cleaved in this rumen in vitro system. These data showed that the modified RUSITEC system is stable, viable and maintained metabolic capacity over a longer period (at least 8 days). This makes many animal experiments obsolete and lead to a significant contribution of the 3R (refine, reduce, replace). The modification of the RUSITEC system enables safe routine use for unlabeled but also for radioactive labelled compounds.

In vitro batch cultures were used to screen four fibrolytic enzyme mixtures at two dosages added to a 60 : 40 silage : concentrate diet containing the C(4) tropical grass Andropogon gayanus grass ensiled at two maturities - vegetative stage (VS) and flowering stage (FS). Based on these studies, one enzyme mixture was selected to treat the same diets and evaluate its impact on fermentation using an artificial rumen (Rusitec). In vitro batch cultures were conducted as a completely randomized design with two runs, four replicates per run and 12 treatments in a factorial arrangement (four enzyme mixtures×three doses). Enzyme additives (E1, E2, E3 and E4) were commercial products and contained a range of endoglucanase, exoglucanase and xylanase activities. Enzymes were added to the complete diet 2 h before incubation at 0, 2 and 4 μl/g of dry matter (DM). Gas production (GP) was measured after 3, 6, 12, 24 and 48 h of incubation. Disappearance of DM (DMD), NDF (NDFD) and ADF (ADFD) were determined after 24 and 48 h. For all four enzyme mixtures, a dosage effect (P>0.05) DM, N, NDF or ADF disappearance after 48 h of incubation nor daily ammonia-N, volatile fatty acids or CH(4) production. However, enzyme application increased (P<0.05) microbial N production in feed particle-associated (loosely-associated) and silage feed particle-bound (firmly associated) fractions. With A. gayanus silage diets, degradation may not be limited by microbial colonization, but rather by the ability of fibrolytic enzymes to degrade plant cell walls within this recalcitrant forage.

A rumen simulation technique was used to evaluate the effects of the complete substitution of a common concentrate mixture (CON) with a mixture consisting solely of by-products from the food industry (BP) at 2 different forage-to-concentrate ratios on ruminal fermentation profile, nutrient degradation, and abundance of rumen microbiota. The experiment was a 2×2 factorial arrangement with 2 concentrate types (CON and BP) and 2 concentrate levels (25 and 50% of diet dry matter). The experiment consisted of 2 experimental runs with 12 fermentation vessels each (n=6 per treatment). Each run lasted for 10d, with data collection on the last 5d. The BP diets had lower starch, but higher neutral detergent fiber (NDF) and fat contents compared with CON. Degradation of crude protein was decreased, but NDF and nonfiber carbohydrate degradation were higher for the BP diets. At the 50% concentrate level, organic matter degradation tended to be lower for BP and CH4 formation per unit of NDF degraded was also lower for BP. The BP mixture led to a higher concentration of propionate and a lower acetate-to-propionate ratio, whereas concentrations of butyrate and caproate decreased. Concentrate type did not affect microbial community composition, except that the abundance of bacteria of the genus Prevotella was higher for BP. Increasing the concentrate level resulted in higher degradation of organic matter and crude protein. At the higher concentrate level, total short-chain fatty acid formation increased and concentrations of isobutyrate and valerate decreased. In addition, at the 50% concentrate level, numbers of protozoa increased, whereas numbers of methanogens, anaerobic fungi, and fibrolytic bacteria decreased. No interaction was noted between the 2 dietary factors on most variables, except that at the higher concentrate level the effects of BP on CH4 and CO2 formation per unit of NDF degraded, crude protein degradation, and the abundance of Prevotella were more prominent. In conclusion, the results of this study suggest that BP in the diet can adequately substitute CON with regard to ruminal fermentation profile and microbiota, showing even favorable fermentation patterns when fed at 50% inclusion rate., (Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

This study investigated the combined impact of essential oil blends (EOBs) and fumaric acid (FA) on ruminal fermentation in dairy cows using the rumen simulation technique (RUSITEC) system. Three rumen-cannulated, non-lactating Holstein Friesian cows served as inoculum donors. The substrate, a total mixed ration (TMR), comprised corn silage, alfalfa hay, and a concentrate mix in a 3:1:1 ratio. The four treatments evaluated were Control (TMR without additives), EFA1 (TMR + EOB1 + FA), EFA2 (TMR + EOB2 + FA), and EFA3 (TMR + EOB3 + FA). Sixteen fermentation chambers were randomly assigned to the treatments, each with four replicates, following a completely randomized design during a 9-day experimental period. EOBs and FA were added at 10 µL/g feed and 3% of TMR, respectively. After a 4-day adaptation, samples were collected for 5 days. Results revealed that EFA1 significantly reduced (p = 0.0351) CH4 emissions by 60.2% without negatively impacting dry matter disappearance, fiber fraction digestibility, pH, or gas volume. All EFAs increased (p < 0.001) the propionate molar proportion and decreased (p < 0.001) the acetate-to-propionate ratio. EFA2 decreased (p < 0.05) the acetate proportion by 3.3% compared to the control. In conclusion, EFA1 is recommended as an effective nutritional intervention to mitigate CH4 emissions and optimize ruminal fermentation in dairy cows. [ABSTRACT FROM AUTHOR]

The effect of cicer milkvetch (Astragalus cicer L.) and sainfoin (Onobrychis viciifolia Scop.) on ruminal fermentation, methane production, and microbial nitrogen synthesis was assessed in two experiments. Experiment 1 analyzed two legumes, cicer milkvetch and sainfoin at two stages (vegetative and late flower) incubated with alfalfa (Medicago sativa) at five inclusion rates 0:100; 25:75, 50:50, 75:25, and 100:0 (as DM) in batch culture. Experiment 2 analyzed vegetative cicer milkvetch and alfalfa incubated in ratios of 25:75, 50:50, 75:25, and 100:0 (as DM) in continuous culture systems (RUSITEC). In batch culture, increased dry matter disappearance (DMD), and propionate percentage (%total), and reduced methane (mg·g−1 DMD) occurred with vegetative cicer milkvetch inclusion. In RUSITEC, DMD linearly increased (P < 0.01), acetate:propionate ratio quadratically decreased, while ammonia (NH3-N) concentration (P < 0.01) and butyrate percentage (%total) linearly decreased (P < 0.05) with increasing inclusion of cicer milkvetch. No differences were observed for methane (CH4) production (mg·g−1 DMD), or short chain fatty acid (SCFA) production (mmol·day−1). Microbial nitrogen synthesis and efficiency of protein synthesis linearly increased (P < 0.05) with increased inclusion of cicer milkvetch. Results suggest cicer milkvetch may result in synchronicity of energy and nitrogen during rumen fermentation, which could enhance cattle production. [ABSTRACT FROM AUTHOR]

This study investigated the combined impact of essential oil blends (EOBs) and fumaric acid (FA) on ruminal fermentation in dairy cows using the rumen simulation technique (RUSITEC) system. Three rumen-cannulated, non-lactating Holstein Friesian cows served as inoculum donors. The substrate, a total mixed ration (TMR), comprised corn silage, alfalfa hay, and a concentrate mix in a 3:1:1 ratio. The four treatments evaluated were Control (TMR without additives), EFA1 (TMR + EOB1 + FA), EFA2 (TMR + EOB2 + FA), and EFA3 (TMR + EOB3 + FA). Sixteen fermentation chambers were randomly assigned to the treatments, each with four replicates, following a completely randomized design during a 9-day experimental period. EOBs and FA were added at 10 µL/g feed and 3% of TMR, respectively. After a 4-day adaptation, samples were collected for 5 days. Results revealed that EFA1 significantly reduced (p = 0.0351) CH4 emissions by 60.2% without negatively impacting dry matter disappearance, fiber fraction digestibility, pH, or gas volume. All EFAs increased (p < 0.001) the propionate molar proportion and decreased (p < 0.001) the acetate-to-propionate ratio. EFA2 decreased (p < 0.05) the acetate proportion by 3.3% compared to the control. In conclusion, EFA1 is recommended as an effective nutritional intervention to mitigate CH4 emissions and optimize ruminal fermentation in dairy cows.

The aim of this study was to distinguish effects due to diet composition from those triggered by ruminal pH on fermentation patterns and microbial profiles in a continuous culture system (RUSITEC). The study followed a 2 × 2 factorial design, with two diets varying in the proportions of forage and concentrate and two pH levels in the culture medium. RUSITEC fermenters were used to simulate rumen fermentation and feed digestibility, fermentation end-products, microbial protein synthesis, microbial community, and long-chain fatty acid profiles in the digesta were determined. Multivariate analyses were applied to summarize the overall results. High concentrate (34% cereal grain, 32% hay) diets were more digestible (p < 0.05) than high forage (10% cereal grain, 78% hay) diets, resulting in a greater (p < 0.05) formation of most fermentation end-products and microbial protein in the rumen. However, there were no significant (p > 0.05) differences between diets in methane production. Ciliate protozoa, anaerobic fungi, some fibrolytic bacteria, hydrogenation of oleic acid, and relative proportion of conjugated linoleic acid were increased (p < 0.05) with high forage diets. A decline in rumen pH from 6.8 to 6.4 decreased (p < 0.05) feed digestibility, protein degradability, and the daily outputs of some fermentation end-products (gas, VFA, acetate, ammonia) but had no effect (p > 0.05) on the synthesis of microbial protein, and on the output of methane, propionate, butyrate or lactate. Minor changes in microbial community profile or the fatty acid relative proportions were observed within this pH range. The overall multivariate analysis revealed a clear discrimination between high-concentrate and high-forage diets, with subtler and less-defined pH effects on ruminal fermentation and microbial communities. [ABSTRACT FROM AUTHOR]

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Background: Brewers' spent grain (BSG) typically contains 20% – 29% crude protein (CP) with high concentrations of glutamine, proline and hydrophobic and non-polar amino acid, making it an ideal material for producing value-added products like bioactive peptides which have antioxidant properties. For this study, protein was extracted from BSG, hydrolyzed with 1% alcalase and flavourzyme, with the generated protein hydrolysates (AlcH and FlaH) showing antioxidant activities. This study evaluated the effects of AlcH and FlaH on gas production, ruminal fermentation characteristics, nutrient disappearance, microbial protein synthesis and microbial community using an artificial rumen system (RUSITEC) fed a high-grain diet. Results: As compared to the control of grain only, supplementation of FlaH decreased (P < 0.01) disappearances of dry matter (DM), organic matter (OM), CP and starch, without affecting fibre disappearances; while AlcH had no effect on nutrient disappearance. Neither AlcH nor FlaH affected gas production or VFA profiles, however they increased (P < 0.01) NH3-N and decreased (P < 0.01) H2 production. Supplementation of FlaH decreased (P < 0.01) the percentage of CH4 in total gas and dissolved-CH4 (dCH4) in dissolved gas. Addition of monensin reduced (P < 0.01) disappearance of nutrients, improved fermentation efficiency and reduced CH4 and H2 emissions. Total microbial nitrogen production was decreased (P < 0.05) but the proportion of feed particle associated (FPA) bacteria was increased with FlaH and monensin supplementation. Numbers of OTUs and Shannon diversity indices of FPA microbial community were unaffected by AlcH and FlaH; whereas both indices were reduced (P < 0.05) by monensin. Taxonomic analysis revealed no effect of AlcH and FlaH on the relative abundance (RA) of bacteria at phylum level, whereas monensin reduced (P < 0.05) the RA of Firmicutes and Bacteroidetes and enhanced Proteobacteria. Supplementation of FlaH enhanced (P < 0.05) the RA of genus Prevotella, reduced Selenomonas, Shuttleworthia, Bifidobacterium and Dialister as compared to control; monensin reduced (P < 0.05) RA of genus Prevotella but enhaced Succinivibrio. Conclusions: The supplementation of FlaH in high-grain diets may potentially protect CP and starch from ruminal degradation, without adversely affecting fibre degradation and VFA profiles. It also showed promising effects on reducing CH4 production by suppressing H2 production. Protein enzymatic hydrolysates from BSG using flavourzyme showed potential application to high value-added bio-products. [ABSTRACT FROM AUTHOR]

The effects of rice bran and corn on ensiling characteristics of potato vines were investigated. Ruminal fermentation of potato vine silage was measured using a rumen simulation technique (Rusitec). Potato vines were treated (fresh weight basis) for ensiling without additives (control), with 15% rice bran and with 15% rice bran plus 3% corn, 5% corn, or 7% corn, respectively. Silage with 5% corn was chosen for a ruminal fermentation trial, based on the results of ensiling quality. Comparison of rumen fermentation between two silage (silage with 5% corn and whole corn silage) diets was conducted using a Rusitec apparatus. Adding rice bran and corn improved the potato vine silage quality (P <.05). Compared with whole corn silage, potato vine silage diet had lower (P <.05) methane production, ruminal pH, acetate, and acetate to propionate ratio and higher (P <.05) total volatile fatty acids, total nitrogen (N), ammonia-N, and microbial-N. These findings indicated that rice bran and corn improved ensiling fermentation characteristics of potato vines, and that additive-treated potato vine silage had no adverse effects on rumen fermentation and could improve the energy and N utilisation and alleviate the methane pollution of the rumen compared with whole corn silage. [ABSTRACT FROM AUTHOR]

This study investigated the effect of treatment of wheat straw using ammonia fiber expansion (AFEX) and exogenous fibrolytic enzymes (Viscozyme) on fiber digestibility, rumen fermentation, microbial protein synthesis, and microbial populations in an artificial rumen system [Rumen Simulation Technique (RUSITEC)]. Four treatments were assigned to 16 vessels (4 per treatment) in 2 RUSITEC apparatuses in a randomized block design. Treatments were arranged as a 2 × 2 factorial using untreated or AFEX-treated wheat straw with or without exogenous fibrolytic enzymes [0 or 500 μg of protein/g straw dry matter (DM)]. Fibrolytic enzymes were applied to straw, prior to sealing in nylon bags. The concentrate mixture was provided in a separate bag within each fermentation vessel. The RUSITECs were adapted for 8 d and disappearance of DM, neutral detergent fiber (NDF), acid detergent fiber (ADF), and crude protein (CP) was measured after 48 h of incubation. Ammonia fiber expansion increased (P < 0.01) the disappearance of wheat straw DM (69.6 vs. 38.3%), NDF (65.6 vs. 36.8%), ADF (61.4 vs. 36.0%), and CP (68.3 vs. 24.0%). Total dietary DM, organic matter (OM), and NDF disappearance was also increased (P ≤ 0.05) by enzymes. Total microbial protein production was greater (P < 0.01) for AFEX-treated (72.9 mg/d) than untreated straw (63.1 mg/d). Total gas and methane (CH4) production (P < 0.01) were also greater for AFEX-treated wheat straw than untreated straw, with a tendency for total gas to increase (P = 0.06) with enzymes. Ammonia fiber expansion increased (P < 0.01) total volatile fatty acid (VFA) production and the molar proportion of propionate, while it decreased (P < 0.01) acetate and the acetate-to-propionate ratio. The AFEX-treated straw had lower relative quantities of fungi, methanogens, and Fibrobacter succinogenes (P < 0.01) and fewer protozoa (P < 0.01) compared to untreated straw. The pH of fermenters fed AFEX-treated straw was lower (P < 0.01) than those fed untreated straw. Both AFEX (P < 0.01) and enzymes (P = 0.02) decreased xylanase activity. There was an enzyme × straw interaction (P = 0.02) for endoglucanase activity. Enzymes increased endoglucanase activity of AFEX-treated wheat straw, but had no effect on untreated straw. The addition of enzymes lowered the relative abundance of Ruminococcus flavefaciens , but increased F. succinogenes. These results indicate that AFEX increased the ruminal disappearance of wheat straw and improved fermentation and microbial protein synthesis in the RUSITEC. [ABSTRACT FROM AUTHOR]

This study investigated the effect of treatment of wheat straw using ammonia fiber expansion (AFEX) and exogenous fibrolytic enzymes (Viscozyme) on fiber digestibility, rumen fermentation, microbial protein synthesis, and microbial populations in an artificial rumen system [Rumen Simulation Technique (RUSITEC)]. Four treatments were assigned to 16 vessels (4 per treatment) in 2 RUSITEC apparatuses in a randomized block design. Treatments were arranged as a 2 × 2 factorial using untreated or AFEX-treated wheat straw with or without exogenous fibrolytic enzymes [0 or 500 μg of protein/g straw dry matter (DM)]. Fibrolytic enzymes were applied to straw, prior to sealing in nylon bags. The concentrate mixture was provided in a separate bag within each fermentation vessel. The RUSITECs were adapted for 8 d and disappearance of DM, neutral detergent fiber (NDF), acid detergent fiber (ADF), and crude protein (CP) was measured after 48 h of incubation. Ammonia fiber expansion increased (P < 0.01) the disappearance of wheat straw DM (69.6 vs. 38.3%), NDF (65.6 vs. 36.8%), ADF (61.4 vs. 36.0%), and CP (68.3 vs. 24.0%). Total dietary DM, organic matter (OM), and NDF disappearance was also increased (P ≤ 0.05) by enzymes. Total microbial protein production was greater (P < 0.01) for AFEX-treated (72.9 mg/d) than untreated straw (63.1 mg/d). Total gas and methane (CH4) production (P < 0.01) were also greater for AFEX-treated wheat straw than untreated straw, with a tendency for total gas to increase (P = 0.06) with enzymes. Ammonia fiber expansion increased (P < 0.01) total volatile fatty acid (VFA) production and the molar proportion of propionate, while it decreased (P < 0.01) acetate and the acetate-to-propionate ratio. The AFEX-treated straw had lower relative quantities of fungi, methanogens, and Fibrobacter succinogenes (P < 0.01) and fewer protozoa (P < 0.01) compared to untreated straw. The pH of fermenters fed AFEX-treated straw was lower (P < 0.01) than those fed untreated straw. Both AFEX (P < 0.01) and enzymes (P = 0.02) decreased xylanase activity. There was an enzyme × straw interaction (P = 0.02) for endoglucanase activity. Enzymes increased endoglucanase activity of AFEX-treated wheat straw, but had no effect on untreated straw. The addition of enzymes lowered the relative abundance of Ruminococcus flavefaciens , but increased F. succinogenes. These results indicate that AFEX increased the ruminal disappearance of wheat straw and improved fermentation and microbial protein synthesis in the RUSITEC. [ABSTRACT FROM AUTHOR]

The reduction of methane emissions by ruminants is a highly desirable goal to mitigate greenhouse gas emissions. Various feed additives have already been tested for their ability to decrease methane production; however, practical use is often limited due to negative effects on rumen fermentation or high costs. Organosulphur compounds from garlic (Allium sativum) and flavonoids have been identified as promising plant-derived compounds which are able to reduce methane production. Here, we evaluated the effects of a combination of garlic powder and bitter orange (Citrus aurantium) extracts, Mootral, on ruminal methane production, ruminal fermentation and the community of methanogenic Archaea by using the rumen simulation technique as ex vivo model. The experiment consisted of an equilibration period of 7 days, an experimental period of 8 days and a withdrawal period of 4 days. During the experimental period three fermenters each were either treated as controls (CON), received a low dose of Mootral (LD), a high dose of Mootral (HD), or monensin (MON) as positive control. Application of Mootral strongly reduced the proportion of methane in the fermentation gas and the production rate of methane. Moreover, the experimental mixture induced a dosedependent increase in the production rate of short chain fatty acids and in the molar proportion of butyrate. Some effects persisted during the withdrawal period. Both, single strand conformation polymorphism and Illumina MiSeq 16S rRNA amplicon sequencing indicated an archaeal community distinct from CON and MON samples in the LD and HD samples. Among archaeal families the percentage of Methanobacteriaceae was reduced during application of both doses of Mootral. Moreover, several significant differences were observed on OTU level among treatment groups and after withdrawal of the additives for LD and HD group. At day 14, 4 OTUs were positively correlated with methane production. In conclusion this mixture of garlic and citrus compounds appears to effectively reduce methane production by alteration of the archaeal community without exhibiting negative side effects on rumen fermentation. [ABSTRACT FROM AUTHOR]

Compared to grass silage (GS)-, corn silage (CS)-based diets appear to increase the efficiency of microbial protein synthesis (EMPS) in the rumen. Opposite results for the EMPS obtained in vitro have raised the question of whether an inadequate supply of amino N for microbes might explain the low EMPS. We examined the effects of supplementation with different N sources in CS on the EMPS and microbial populations in vitro . GS and CS were used as substrates for in vitro incubation. CS was non-supplemented or supplemented with urea, mixed amino acids (AA), peptone, or protein to adjust the N content to that of GS. Degradation of organic matter (OM) and crude protein (CP) revealed a positive effect of all N supplements, except protein. Additionally, N supplementation increased fiber degradation of CS. Peptone primarily stimulated hemicellulolytic activity and urea stimulated cellulolytic activity. The EMPS of CS was improved by all N supplements, with peptone and urea exhibiting the highest increase (57% and 54%, respectively), followed by AA mix (40%) and protein (11%) compared to that of CS alone (111 g microbial CP kg −1 fermented OM). However, the level of EMPS detected with GS (200 g microbial CP kg −1 fermented OM) was not achieved. Protozoal 18S rRNA gene copy numbers were negatively correlated with the EMPS, whereas no correlation was found between total bacteria and the EMPS. A stimulating effect of urea, AA mix, and peptone was detected for Ruminococcus albus and Prevotella bryantii , whereas Fibrobacter succinogenes was inhibited by N supplementation. This indicated that neither the amount of available N nor the N source was the only limiting factor in the low EMPS values of CS in vitro . Information is also provided on the stimulating effects of different N sources on several microbial species in mixed rumen culture. [ABSTRACT FROM AUTHOR]

Abstract: BACKGROUND: Molassed sugar beet pulp (Bp) is a viable alternative to grains in cattle nutrition for reducing human edible energy input. Yet little is known about the effects of high inclusion rates of Bp on rumen microbiota. This study used an in vitro approach and the quantitative polymerase chain reaction technique to establish the effects of a graded replacement of maize grain (MG) by Bp on the ruminal microbial community, fermentation profile and nutrient degradation. RESULTS: Six different amounts of Bp (0–400 g kg−1), which replaced MG in the diet, were tested using the in vitro semi‐continuous rumen simulation technique. The increased inclusion of Bp resulted in greater dietary content and degradation of neutral detergent fibre (P < 0.01). Further, Bp feeding enhanced (P < 0.01) the abundance of genus Prevotella and shifted (P < 0.01) the short‐chain fatty acid patterns in favour of acetate and propionate and at the expense of butyrate. A total replacement of MG with Bp resulted in an increased daily methane production (P < 0.01). CONCLUSION: Results suggest positive effects of the replacement of MG by Bp especially in terms of stimulating ruminal acetate and propionate fermentation. However, high replacement rates of Bp resulted in lowered utilization of ammonia and higher ruminal methane production. © 2017 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

In vitro fermentation systems such as the rumen simulation technique (RUSITEC) are frequently used to assess dietary manipulations in livestock, thereby limiting the use of live animals. Despite being in use for nearly 40 years, improvements are continually sought in these systems to better reflect and mimic natural processes in ruminants. The aim of this study was to evaluate the effect of forage preparation, i.e., frozen minced (FM) and freeze-dried and ground (FDG), on the ruminal microbiota and on fermentation characteristics when included as a substrate in a RUSITEC system. A completely randomized design experiment was performed over a 15-day period, with 7 days of adaptation and an 8-day experimental period. Fermentation parameters (total gas, CH4, and volatile fatty acid production) were analyzed on a daily basis over the experimental period and the archaeal and bacterial microbiota (liquid-associated microbes [LAM] and solid-associated microbes [SAM] was assessed at 0, 5, 10, and 15 days using high-throughput sequencing of the 16S rRNA gene. Results from this study suggested a tendency (P = 0.09) of FM treatment to increase daily CH4 (mg/d) production by 16.7% when compared with FDG treatment. Of the major volatile fatty acids (acetate, propionate, and butyrate), only butyrate production was greater (P = 0.01) with FM treatment compared with FDG substrate. The archaeal and bacterial diversity and richness did not differ between the forage preparations, although feed particle size of the forage had a significant effect on microbial community structure in the SAM and LAM samples. The Bacteroidetes phylum was more relatively abundant in the FM substrate treatment, while Proteobacteria was enriched in the FDG treatment. At the genus-level, Butyrivibrio, Prevotella, and Roseburia were enriched in the FM substrate treatment and Campylobacter and Lactobacillus in the FDG substrate treatment. Evidence from this study suggests that forage preparation affects CH4 production, butyrate production, and the structure of the rumen microbiota during in vitro fermentation.

In vitro fermentation systems such as the rumen simulation technique (RUSITEC) are frequently used to assess dietary manipulations in livestock, thereby limiting the use of live animals. Despite being in use for nearly 40 years, improvements are continually sought in these systems to better reflect and mimic natural processes in ruminants. The aim of this study was to evaluate the effect of forage preparation, i.e., frozen minced (FM) and freeze-dried and ground (FDG), on the ruminal microbiota and on fermentation characteristics when included as a substrate in a RUSITEC system. A completely randomized design experiment was performed over a 15-day period, with 7 days of adaptation and an 8-day experimental period. Fermentation parameters (total gas, CH4, and volatile fatty acid production) were analyzed on a daily basis over the experimental period and the archaeal and bacterial microbiota (liquid-associated microbes [LAM] and solid-associated microbes [SAM] was assessed at 0, 5, 10, and 15 days using high-throughput sequencing of the 16S rRNA gene. Results from this study suggested a tendency (P = 0.09) of FM treatment to increase daily CH4 (mg/d) production by 16.7% when compared with FDG treatment. Of the major volatile fatty acids (acetate, propionate, and butyrate), only butyrate production was greater (P = 0.01) with FM treatment compared with FDG substrate. The archaeal and bacterial diversity and richness did not differ between the forage preparations, although feed particle size of the forage had a significant effect on microbial community structure in the SAM and LAM samples. The Bacteroidetes phylum was more relatively abundant in the FM substrate treatment, while Proteobacteria was enriched in the FDG treatment. At the genus-level, Butyrivibrio, Prevotella, and Roseburia were enriched in the FM substrate treatment and Campylobacter and Lactobacillus in the FDG substrate treatment. Evidence from this study suggests that forage preparation affects CH4 production, butyrate production, and the structure of the rumen microbiota during in vitro fermentation. [ABSTRACT FROM AUTHOR]

Rumen microbiota have important metabolic functions for the host animal. This study aimed at characterizing changes in rumen microbial abundances and fermentation profiles using a severe subacute ruminal acidosis (SARA) in vitro model, and to evaluate a potential modulatory role of plant derived alkaloids (PDA), containing quaternary benzophenanthridine and protopine alkaloids, of which sanguinarine and chelerythrine were the major bioactive compounds. Induction of severe SARA strongly affected the rumen microbial composition and fermentation variables without suppressing the abundance of total bacteria. Protozoa and fungi were more sensitive to the low ruminal pH condition than bacteria. Induction of severe SARA clearly depressed degradation of fiber (P < 0.001), which came along with a decreased relative abundance of fibrolytic Ruminococcus albus and Fibrobacter succinogenes (P < 0.001). Under severe SARA conditions, the genus Prevotella, Lactobacillus group, Megasphaera elsdenii, and Entodinium spp. (P < 0.001) were more abundant, whereas Ruminobacter amylophilus was less abundant. SARA largely suppressed methane formation (−70%, P < 0.001), although total methanogenic 16S rRNA gene abundance was not affected. According to principal component analysis, Methanobrevibacter spp. correlated to methane concentration. Addition of PDA modulated ruminal fermentation under normal conditions such as enhanced (P < 0.05) concentration of total SCFA, propionate and valerate, and increased (P < 0.05) degradation of crude protein compared with the unsupplemented control diet. Our results indicate strong shifts in the microbial community during severe SARA compared to normal conditions. Supplementation of PDA positively modulates ruminal fermentation under normal ruminal pH conditions. [ABSTRACT FROM AUTHOR]

Copyright of Canadian Journal of Animal Science is the property of Canadian Science Publishing and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

A rumen simulation technique (Rusitec) apparatus with eight 800 ml fermentation vessels was used to investigate the effects of rumen degradable protein (RDP) level and non-fibre carbohydrate (NFC) type on ruminal fermentation, microbial growth, and populations of ruminal cellulolytic bacteria. Treatments consisted of two NFC types (starch and pectin) supplemented with 0 g/d (low RDP) or 1.56 g/d (high RDP) sodium caseinate. Apparent disappearance of dry matter and organic matter was greater for pectin than for starch treatment (P<0.01) with low or high RDP. A NFC × RDP interaction was observed for neutral detergent fibre disappearance (P=0.01), which was lower for pectin than for starch only under low RDP conditions. Compared with starch, pectin treatment increased the copy numbers of Ruminococcus albus (P≤0.01) and Ruminococcus flavefaciens (P≤0.09), the molar proportion of acetate (P<0.01), the acetate:propionate ratio (P<0.01), and methane production (P<0.01), but reduced the propionate proportion (P<0.01). Increasing dietary RDP increased the production of total VFA (P=0.01), methane (P<0.01), ammonia N (P<0.01), and microbial N (P<0.01). Significant NFC × RDP interaction and interaction tendency were observed for ammonia N production (P=0.01) and daily N flow of total microorganisms (P=0.07), which did not differ under low RDP conditions, but pectin produced greater microbial N and less ammonia N than starch with increased RDP. Results showed NFC type, RDP level, and their interaction affected ruminal fermentation and microbial growth, and under sufficient ruminal degradable N pectin had greater advantage in microbial N synthesis than starch in vitro. [ABSTRACT FROM AUTHOR]

Copyright of Canadian Journal of Animal Science is the property of Canadian Science Publishing and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Canadian Journal of Animal Science is the property of Canadian Science Publishing and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Abstract: Eremophila glabra Juss. (Scrophulariaceae), a native Australian shrub, has been demonstrated to have low methanogenic potential in a batch in vitro fermentation system. The present study aimed to test longer-term effects of E. glabra on rumen fermentation characteristics, particularly methane production and the methanogen population, when included as a component of a fermentation substrate in an in vitro continuous culture system (Rusitec). E. glabra was included at 150, 250, 400 g/kg DM (EG15, EG25, and EG40) with an oaten chaff and lupin-based substrate (control). Overall, the experiment lasted 33 days, with 12 days of acclimatization, followed by two periods during which fermentation characteristics (total gas, methane and VFA productions, dry matter disappearance, pH) were measured. The number of copies of genes specifically associated with total bacteria and cellulolytic bacteria (16S rRNA gene) and total ruminal methanogenic archaeal organisms (the methyl coenzyme M reductase A gene (mcrA)) was also measured during this time using quantitative real-time PCR. Total gas production, methane and volatile fatty acid concentrations were significantly reduced with addition of E. glabra. At the end of the experiment, the overall methane reduction was 32% and 45% for EG15 and EG25 respectively, compared to the control, and the reduction was in a dose-dependent manner. Total bacterial numbers did not change, but the total methanogen population decreased by up to 42.1% (EG40) when compared to the control substrate. The Fibrobacter succinogenes population was reduced at all levels of E. glabra, while Ruminococcus albus was reduced only by EG40. Our results indicate that replacing a portion of a fibrous substrate with E. glabra maintained a significant reduction in methane production and methanogen populations over three weeks in vitro, with some minor inhibition on overall fermentation at the lower inclusion levels. [Copyright &y& Elsevier]

A rumen simulation technique apparatus with eight 800 mL fermentation vessels was used to investigate the effects of rumen degradable protein (RDP) level and non-fibre carbohydrate (NFC) type on ruminal fermentation, microbial growth, and populations of ruminal cellulolytic bacteria. Treatments consisted of two NFC types (starch and inulin) supplemented with 0 g/d (low RDP) or 1.56 g/d (high RDP) sodium caseinate. No significant differences existed among dietary treatments in the apparent disappearance of dietary nutrients except for dietary N, which increased with increased dietary RDP (P<0.001). Compared with starch, inulin treatments reduced the molar proportion of acetate (P<0.001), the acetate:propionate ratio (P<0.001), and methane production (P=0.006), but increased the butyrate proportion (P<0.001). Increased dietary RDP led to increases in production of total volatile fatty acid (P=0.014) and methane (P=0.050), various measures of N (P⩽0.046), and 16s rDNA copy numbers of Ruminococcus flavefaciens (P⩽0.010). Non-fibre carbohydrate source did not affect daily microbial N flow regardless of dietary RDP, but ammonia N production was lower for inulin than for starch treatments under high RDP conditions (P<0.001). Compared with starch treatments, inulin depressed the copy numbers of Fibrobacter succinogenes in solid fraction (P=0.023) and R. flavefaciens in liquid (P=0.017) and solid fractions (P=0.007), but it increased the carboxymethylcellulase activity in solid fraction (P=0.045). Current results suggest that starch and inulin differ in ruminal volatile fatty acid fermentation but have similar effects on ruminal digestion and microbial synthesis in vitro, although inulin suppressed the growth of partial ruminal cellulolytic bacteria. [ABSTRACT FROM AUTHOR]

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 r DNA 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. [ABSTRACT FROM AUTHOR]

An in vitro experiment was designed to investigate the effects of incubating two forages with a different energy/nitrogen (N) ratio [perennial ryegrass (GR) vs red clover (RC)] on the efficiency of N utilisation by rumen microbes. Second-cut forages were incubated in artificial rumen fermenters (n = 8). Ryegrass represented a supply of quickly available N and energy for the rumen microorganism which led to a rapid fermentation and bacterial growth 2-4 h after feeding. Ryegrass also promoted greater numbers of anaerobic fungi, methanogens and cellulolytic bacteria, which tended to increase neutral detergent fibre disappearance, gas production, volatile fatty acid and methane production than observed using RC diets. On the contrary, RC provided slowly degradable N and energy, which led to a relatively slow bacterial growth (4-8 h after feeding). In terms of diet utilisation, RC diets promoted a higher N outflow (mainly as undegraded-N) and efficiency of microbial protein synthesis per organic matter disappeared. Even so, microbial protein yield was similar on both diets indicating a better N capture by microorganisms fed GR than in those fed RC diets. The use of 15N-labelled forages demonstrated that this high ammonia incorporation by bacteria-fed GR occurred mainly during the early fermentation coinciding with the highest bacterial growth. In conclusion, this experiment demonstrated that the use of isotopic labelling combined with molecular techniques provided an insight into forage utilisation by the rumen microbes; GR diets led to a better efficiency of N utilisation compared with RC; moreover the lower N outflow on GR diets may be partially compensated for a higher proportion of microbial protein leaving the system and the greater volatile fatty acid production. These findings seem to indicate that RC grazing may increase the N pollution compared with GR without substantial improvements on the rumen function, however this must be confirmed in vivo. Balancing energy and nitrogen in the rumen is essential for the optimisation of ruminant diets. Our in vitro findings indicated that red clover provides an excess of rumen-degradable nitrogen, which leads to a lower efficiency of nitrogen utilisation by the rumen microorganisms than observed in ryegrass diets. As a result, red clover grazing may increase nitrogen pollution compared with ryegrass without substantial improvement to rumen function. [ABSTRACT FROM AUTHOR]

Roots of rhubarb (Rheum spp.) and bark of alder buckthorn (Frangula alnus) were tested as feed additives for decreasing ruminal methane production released from anaerobic fermentation of a forage-based diet in a rumen-simulating fermenter (Rusitec). Sixteen fermentation units (vessels) were set up for the experiment lasting 19 d. Treated vessels were supplied with 1 g/d of rhubarb or alder buckthorn (4 vessels per plant species); another 4 vessels received 12 μM sodium monensin (positive control), and the remaining 4 vessels were controls (no additive). Upon termination of the experimental period, batch cultures were inoculated with the liquid contents of the vessels for examining in vitro fermentation kinetics of cellulose, starch, barley straw, and the same substrate used in the Rusitec cultures. Monensin induced changes in fermentation in agreement with those reported in the literature, and inocula from those cultures decreased the fermentation rate and total gas produced in the gas kinetics study. Rhubarb decreased methane production, associated with limited changes in the profile of volatile fatty acids throughout the duration of the study, whereas digestibility and total volatile fatty acids production were not affected. Rhubarb inocula did not affect gas production kinetics except for cellulose. Alder buckthorn decreased only methane concentration in fermentation gas, and this effect was not always significant. The use of rhubarb(milled rhizomes of Rheum spp.) in the diets of ruminants may effectively modulate ruminal fermentation by abating methane production, thus potentially involving productive and environmental benefits. [ABSTRACT FROM AUTHOR]

Four ruminally and duodenally cannulated sheep and 8 Rusitec fermenters were used to determine the effects of forage to concentrate (F:C) ratio and type of forage in the diet on ruminal fermentation and microbial protein synthesis. The purpose of the study was to assess how closely fermenters can mimic the dietary differences found in vivo. The 4 experimental diets contained F:C ratios of 70:30 or 30:70 with either alfalfa hay or grass hay as the forage. Microbial growth was determined in both systems using 15N as a microbial marker. Rusitec fermenters detected differences between diets similar to those observed in sheep by changing F:C ratio on pH; neutral detergent fiber digestibility; total volatile fatty acid concentrations; molar proportions of acetate, propionate, butyrate, isovalerate, and caproate; and amylase activity. In contrast, Rusitec fermenters did not reproduce the dietary differences found in sheep for NH3-N and lactate concentrations, dry matter (DM) digestibility, proportions of isobutyrate and valerate, carboxymethylcellulase and xylanase activities, and microbial growth and its efficiency. Regarding the effect of the type of forage in the diet, Rusitec fermenters detected differences between diets similar to those found in sheep for most determined parameters, with the exception of pH, DM digestibility, butyrate proportion, and carboxymethylcellulase activity. Minimum pH and maximal volatile fatty acid concentrations were reached at 2 h and at 6 to 8 h postfeeding in sheep and fermenters, respectively, indicating that feed fermentation was slower in fermenters compared with that in sheep. There were differences between systems in the magnitude of most determined parameters. In general, fermenters showed lower lactate concentrations, neutral detergent fiber digestibility, acetate:propionate ratios, and enzymatic activities. On the contrary, fermenters showed greater NH3-N concentrations, DM digestibility, and proportions of propionate, butyrate, isovalerate, valerate, and caproate. Values of efficiency of microbial growth were greater in fermenters compared with sheep for 70:30 diets, but they were lower for 30:70 diets. Differences between fermentation in sheep and fermenters can be mainly attributed to the lack of absorption in fermenters, differences in solid retention time, and compartmentalization in the Rusitec system. In general, the Rusitec system simulated more closely the in vivo fermentation of high-forage diets compared with high-concentrate diets. [ABSTRACT FROM AUTHOR]

Four ruminally and duodenally cannulated sheep and 8 Rusitec fermenters were used to determine the effects of dietary characteristics on microbial populations and bacterial diversity. The purpose of the study was to assess how closely fermenters can mimic the differences between diets found in vivo. The 4 experimental diets contained forage to concentrate (F:C) ratios of 70:30 (high forage; HF) or 30:70 (high concentrate; HC) with either alfalfa hay (A) or grass hay (G) as the forage. Total bacterial numbers were greater in the rumen of sheep fed HF diets compared with those fed HC diets, whereas the opposite was found in fermenters. The numbers of cellulolytic bacteria were not affected by F:C ratio in any fermentation system, but cellulolytic numbers were 2.7 and 1.8 times greater in sheep than infermenters for HF and HC diets, respectively. Neither total bacterial nor cellulolytic numbers were affectedby the type of forage in sheep or fermenters. Decreasing F:C ratio increased total protozoa and Entodiniae numbers in sheep by about 29 and 25%, respectively, but it had no effect in fermenters. Isotrichidae and Ophryoscolecinae numbers in sheep were not affected by changing F:C ratio, but both disappeared completely from fermenters fed HC diets. Total protozoa and Entodiniae numbers were greater in sheep fed A diets than in those fed G diets, whereas the opposite was found in fermenters. Results indicate that under the conditions of the present study, protozoa population in Rusitec fermenters was not representative of that in the rumen of sheep fed the same diets. In addition, protozoa numbers in fermenters were 121 and 226 times lower than those in the sheep rumen for HF and HC diets, respectively. The automated ribosomal intergenic spacer analysis of the 16S ribosomal DNA was used to analyze the diversity of liquid- and solid-associated bacteria in both systems. A total of 170 peaks were detected in the automated ribosomal intergenic spacer analysis electropherograms of bacterial pellets across the full set of 64 samples, from which 160 were detected in at least 1 individual from each system (sheep or fermenter). Diversity of liquid-associated bacterial pellets was greater with G diets in fermenters but seemed to be unaffected by diet in sheep. Bacterial diversity in solid-associated bacteria pellets was greater for G diets compared with A diets in sheep and fermenters. Different conditions in the fermenters compared with sheep rumen might have caused a selection of some bacterial strains. [ABSTRACT FROM AUTHOR]

The goal of this study was to identify dietary factors that affect fermentation and efficiency of microbial crude protein (CPM) synthesis in the rumen in vitro. We used 16 total mixed, dairy cow rations with known digestibilities that varied in ingredient composition and nutrient content. Each ration was incubated in a Rusitec (n = 3) for 15 d, and fermentation of different fractions was assessed. Observed extents of fermentation in 24 h were 35 to 47% for organic matter, 25 to 60% for crude protein, 3 to 28% for neutral detergent fiber, and 31 to 45% for gross energy. Organic matter fermentation depended on the content of crude protein and neutral detergent fiber in the ration. We studied net synthesis of CPM using an 15N dilution technique and found that 7 d of continuous 15N application are needed to achieve an 15N enrichment plateau in the N of isolated microbes in this type of study. The efficiency of CPM synthesis was 141 to 286 g/kg of fermented organic matter or 4.9 to 11.1 g/MJ of metabolizable energy, and these ranges agree with those found in the literature. Multiple regressions to predict the efficiency of CPM synthesis by diet data showed that crude protein was the only dietary chemical fraction that had a significant effect. Fat content and the inclusion rate of corn silage in the ration also tended to improve efficiency. We suggest that microbial need for preformed amino acids may explain the crude protein effect. A large part of the variation in efficiency of microbial activity still remains unexplained. [ABSTRACT FROM AUTHOR]

• Biochar did not affect rumen fermentation and nutrient disappearance. • Biochar had no effect on total gas and methane production. • Biochar did not affect protozoa populations and microbial protein synthesis. • Biochar had no effect on rumen microbiota diversity and richness. This study investigated the effects of including biochars, which differed in post-pyrolysis treatment, on nutrient disappearance, total gas and methane (CH 4) emissions, rumen fermentation, microbial protein synthesis and rumen microbiota in an artificial rumen system (RUSITEC) fed a barley silage-based diet. The basal diet contained (g/kg dry matter (DM) basis); 600 barley silage, 270 barley grain, 100 canola meal and 30 mineral/vitamin supplement. Three spruced-based biochars were treated post-pyrolysis with either zinc chloride, a hydrochloric acid/nitric acid mixture or sulfuric acid and each included at 20 g/kg of diet DM. Treatments were assigned to sixteen fermenters (n = 4/treatment) in two RUSITEC units in a randomized complete block design. The experiment was conducted over 15 d, with 8 d of adaptation and 7 d of sampling and data collection. Nutrient disappearance of DM, organic matter (OM), acid detergent fiber (ADF), nutrient detergent fiber (NDF) and starch were determined after 48 h of incubation from d 9–12, and microbial protein synthesis was measured from d 14−15. Data were analyzed using PROC MIXED in SAS, with fixed effects of treatment and day of sampling (repeated measure) and random effects of RUSITEC unit and fermenters. Biochar inclusion did not affect disappearance of DM (P = 0.49), OM (P = 0.60), CP (P = 0.47), NDF (P = 0.48), ADF (P = 0.11) or starch (P = 0.58). Moreover, biochar inclusion did not affect total gas production (P = 0.31) or CH 4 produced expressed as mg/g of DM incubated (P = 0.74), and mg/g (P = 0.64), or mg/d (P = 0.70) of DM disappeared. Compared to the control, there was a tendency (P = 0.06) for reduced CH 4 production with biochar inclusion when CH 4 was expressed as percent of total gas produced. Biochar inclusion did not affect total VFA (P = 0.56) or NH 3 -N (P = 0.20) production. Microbial protein synthesis was not affected by biochar inclusion (P > 0.05). Total protozoa counts were also unaffected by biochar (P = 0.37) nor did it impact (P > 0.05) the alpha or beta diversity of bacterial populations. In conclusion, the biochars evaluated in this study appeared to have little to no impact on nutrient disappearance, total gas and CH 4 production, rumen fermentation, and rumen microbiota in the RUSITEC system. [ABSTRACT FROM AUTHOR]

Simple Summary: Citrus pulp is the main by-product obtained from citrus processing. The high-moisture content of this by-product makes it rapidly perishable, its accumulation can cause environmental problems, and it causes high disposal costs for citrus processing factories. Therefore, alternative uses for citrus pulp are necessary, its use in ruminant feeding being one of the most feasible ones. In this study, we assessed the effects of replacing extruded maize in a diet for dairy sheep (20% of diet) by dried citrus pulp using an in vitro technique (Rusitec fermenters). Results showed some positive effects of citrus pulp on diet degradability and in vitro fermentation parameters. The growth of ruminal microbes and bacterial diversity were essentially unaffected. Our results indicate that maize in dairy sheep diets can be totally replaced by dried citrus pulp without negatively affecting ruminal fermentation. The use of citrus pulp would reduce the amount of human-edible ingredients used in the diet of dairy sheep. Citrus pulp is a highly abundant by-product of the citrus industry. The aim of this study was to assess the effects of replacing extruded maize (EM; 20% of total diet) by dried citrus pulp (DCP; 20%) in a mixed diet on rumen fermentation and microbial populations in Rusitec fermenters. The two diets contained 50% alfalfa hay and 50% concentrate, and the same protein level. Four Rusitec fermenters were used in a cross-over design with two 13-d incubation runs. After 7-d of diet adaptation, diet disappearance, fermentation parameters, microbial growth, and microbial populations were assessed. Fermenters receiving the DCP showed greater pH values and fiber disappearance (p < 0.001) and lower methane production (p = 0.03) than those fed EM. Replacing EM by DCP caused an increase in the proportions of propionate and butyrate (p < 0.001) and a decrease in acetate (p = 0.04). Microbial growth, bacterial diversity, and the quantity of bacteria and protozoa DNA were not affected by the diet, but the relative abundances of fungi and archaea were greater (p < 0.03) in solid and liquid phases of DCP fermenters, respectively. Results indicate that DCP can substitute EM, promoting a more efficient ruminal fermentation. [ABSTRACT FROM AUTHOR]