Search Results (194)

Efficient utilization of lignocellulosic biomass by the rumen microbiome is critical for improving feed efficiency in ruminants, yet the development of stable, functionally specialized microbial consortia remains limited. This study aimed to assemble substrate-adapted rumen microbial consortia using an ecology-guided enrichment approach. Rumen fluid collected from cannulated Angus × Hereford heifers was sequentially enriched over 10 generations on four substrates with distinct cell wall characteristics: alfalfa, barley straw, carboxymethyl cellulose (CMC), and xylan. Fermentation parameters, including gas production and volatile fatty acids (VFAs), and bacterial community dynamics were analyzed, and selected consortia (alfalfa and xylan) were evaluated for stability following one month of cryopreservation. Across enrichments, total VFA concentrations declined (e.g., xylan: 109.8 mM (G0) to 56.37 mM (G10)), accompanied by reduced gas production and decreased alpha diversity, indicating substrate-driven selection. Distinct functional profiles emerged, including increased propionate in alfalfa consortia, higher acetate in barley straw, lactate–propionate cross-feeding with CMC, and caproate production (6.3 mM at G10) in xylan enrichments associated with Caproiciproducens and Megasphaera. Cryopreserved consortia retained core community structure and fermentation characteristics upon revival. These results demonstrate that substrate-driven enrichment can generate stable, functionally specialized rumen consortia and provide a framework for developing ecologically compatible microbial communities with potential applications in improving rumen fermentation efficiency. Full article

This study evaluated the effects of replacing whole-crop maize silage with varying proportions of Pennisetum giganteum silage on rumen fermentation, microbial composition, and metabolic function in beef cattle. A single-factor completely randomized design was employed using 50 healthy crossbred Simmental cattle aged 11–12 months (average body weight: 251.08 ± 51.54 kg). Animals were randomly assigned to five groups, with 10 replicates per group and one animal per replicate. Diets contained 0% (Group A), 25% (Group B), 50% (Group C), 75% (Group D), or 100% (Group E) Pennisetum giganteum silage replacing whole-crop maize silage over a 67-day feeding period, including a 7-day adaptation phase. Rumen fluid samples were collected via rumen catheter at the end of the trial to assess bacterial diversity and functional characteristics. Increasing the proportion of Pennisetum giganteum silage resulted in quadratic changes in volatile fatty acids (VFAs) and propionate (PA) concentrations (p < 0.05), while ammonia nitrogen (NH₃-N) increased linearly (p < 0.05). No significant differences were observed in α- or β-diversity among groups (p > 0.05). Group C exhibited significantly higher relative abundances of Verrucomicrobiota and Prevotellaceae_UCG_003 compared with the other groups (p < 0.05). At the phylum level, Proteobacteria increased linearly, whereas Spirochaetota decreased linearly; at the genus level, Treponema decreased linearly (p < 0.05). LEfSe analysis indicated enrichment of g__Prevotellaceae_UCG_003 and o__WCHB1_41 in Group C, while the relative abundances of f__Enterobacteriaceae and g__Citrobacter were elevated in Group E. Under the conditions of this study, replacing 50% of whole-crop maize silage with Pennisetum giganteum silage enhanced rumen fermentation efficiency and modulated key microbial populations in beef cattle. Full article

The rumen microbiota plays a key role in nutrient fermentation and short-chain fatty acid (SCFA) production in ruminants. However, the impacts of different feeding regimes on rumen microbial composition, functional potential, and metabolic outputs remain unclear. In this study, rumen fluid samples were collected from 12 Longdong goats (Capra hircus), which were divided into four groups based on feeding regime and coat color: housed white goats (n = 3), housed black goats (n = 3), grazing white goats (n = 3), and grazing black goats (n = 3). Samples were analyzed using high-throughput sequencing combined with functional annotation (KEGG and CAZy) and targeted SCFA profiling. Distinct differences in microbial community composition were observed primarily between feeding regimes, with enrichment of taxa related to carbohydrate degradation and fermentation. Functional analyses revealed significant shifts in metabolic pathways, particularly those associated with carbohydrate metabolism, energy production, and glycan biosynthesis. Several glycoside hydrolase and glycosyltransferase families showed differential abundances across groups. Consistently, SCFA concentrations varied significantly among feeding regimes. Correlation analyses further demonstrated strong associations between key microbial taxa, functional pathways, and specific SCFAs. Overall, these results indicate that feeding regime, rather than coat color, plays a dominant role in shaping rumen microbial structure, functional capacity, and fermentation characteristics, providing insights into microbial mechanisms underlying rumen metabolism and informing feeding strategy optimization. Full article

This study aimed to evaluate the effect of bromoform (CHBr₃) dose on in vitro rumen fermentation and on CHBr₃ and dibromomethane (CH₂Br₂) concentrations in solution and the gas cap. In vitro treatments consisted of CHBr₃ (DOSE: 0, 1, 10, 100, 1000, 10,000 µg of CHBr₃), with five replicates per DOSE at each time-point (TIME: 0, 0.25, 0.5, 1, 2, 3, 4, 5, 6, 12, 24, 48, and 72 h). The 10,000 µg CHBr₃ DOSE inhibited fermentation completely and was removed from the dataset. The acetate:propionate ratio, nitrogen, and methane (CH₄) produced per gram of DMD decreased as DOSE increased (p = 0.01). As the DOSE increased, CH₄ decreased, and H₂ increased in a dose-dependent manner (p < 0.01). The CHBr₃ concentration dropped below the detection limit within 3 h of incubation. Dibromomethane concentrations for DOSE 1000 and 100 µg of CHBr₃ increased in solution and gas cap beginning at 0.25 h and 1 h post incubation and plateaued by hour 3 and 5, respectively (p < 0.01). The addition of CHBr₃ alters the molar proportion of volatile fatty acids, decreases CH₄, and increases H₂ production, and it is dehalogenated to CH₂Br₂ within 3 h of incubation in an in vitro system. Full article

Pasture plants can contribute to ruminant nutrition and may, depending on composition, influence rumen fermentation and methane production. This study evaluated the nutritional composition, bioactive compounds, and methane production potential of 32 terrestrial plant species commonly foraged by goats in Malta. Dried plant samples were analysed for proximate composition using near-infrared spectroscopy, total polyphenols using the Folin–Ciocalteu assay, antioxidant activity using the DPPH assay, and methane production using an in vitro rumen fermentation system incubated for 48 h, with rumen fluid pooled from three goats (analyses performed in triplicate). Crude protein ranged from 1.16 to 31.97% DM, neutral detergent fibre from 12.29 to 48.89%, and ash from 9.69 to 17.20% across species. Total polyphenolic content varied from 0.07 to 1.30% (w/w), while antioxidant activity (IC₅₀) ranged from 0.37 to 55.9 mg/mL. Methane production after 48 h ranged from 30.39 to 198.26 L CH₄ kg⁻¹, indicating variation in fermentation characteristics among species. These results indicate that Rumex bucephalophorus and Urtica pilulifera demonstrated relatively high protein or bioactive values and comparatively lower in vitro methane-related parameters under the conditions tested. Full article

Corn dust is an abundant agro-industrial by-product with potential as an alternative energy source. Its use in animal feeding, however, is restricted by high fiber content and low digestibility. This study evaluated the effects of non-starch polysaccharide (NSP) enzymes and yeast (Candida tropicalis KKU20) on the chemical composition, fermentation characteristics, and microbial populations of fermented corn dust. The experiment was conducted using a completely randomized design with a 3 × 2 factorial arrangement plus an additional control treatment. Factor A consisted of three levels of enzyme supplementation (0.02%, 0.04%, and 0.06% of dry matter), and Factor B consisted of yeast supplementation (without yeast or with C. tropicalis KKU20, approximately 1 × 10¹³ cells/g of inoculum). The control treatment consisted of fermented corn dust without enzyme or yeast supplementation. Samples were fermented for 15 days prior to analysis. Yeast inoculation increased crude protein and non-fiber carbohydrate contents while reducing neutral detergent fiber, acid detergent fiber, and acid detergent lignin (p < 0.05). Significant enzyme × yeast interactions were observed for several components, particularly fiber fractions (p < 0.05). The reduction in fiber was more pronounced when enzymes were combined with yeast. Predicted energy values, including metabolizable and digestible energy, were increased following yeast supplementation (p < 0.05). Fermentation characteristics were mainly affected by yeast. Yeast-treated samples exhibited higher pH and ammonia–nitrogen concentrations, indicating increased nitrogen turnover during fermentation. In contrast, lactic and propionic acid concentrations were higher in treatments without yeast, while yeast inoculation was associated with lower acetic acid and slightly higher butyric acid levels. Microbial analysis indicated interactions between treatments for lactic acid bacteria populations, reflecting competition for available substrates. No coliform bacteria were detected, indicating acceptable hygienic quality. Overall, yeast inoculation modified the chemical composition of corn dust, particularly by increasing crude protein and reducing fiber fractions, while NSP enzymes contributed to fiber degradation, especially when combined with yeast. However, these changes reflect compositional modification rather than confirmed feeding value, and further evaluation under rumen or in vivo conditions is required. Full article

A study was conducted to evaluate the effect of neutral detergent fiber (NDF) levels of calf starter and weaning time on growth, rumen fermentation characteristics, serum metabolites, and rumen microbial diversity of Holstein calves. A total of 24 newly born male Holstein calves were randomly distributed to four groups in a completely randomized design with a 2 × 2 factorial arrangement of NDF levels (14% and 24%) and weaning time (d 44 and d 54). There was no interaction between starter NDF levels and weaning time for any trait except rumen acetic acid in the immediate post-weaning phase (p = 0.013). Starter NDF levels had no effect on growth, feed intake, and hay intake. Late-weaned calves had greater (p = 0.050) weight gain in the pre-weaning phase whereas, early-weaned calves showed greater weight gain (p = 0.004) and starter intake (p = 0.004) in the post-weaning phase although overall weight gain, and starter and hay intakes were not affected by weaning time. Rumen pH, ammonia nitrogen, and most volatile fatty acids remained unaffected by starter NDF levels and weaning except isobutyric acid which was greater in calves fed 24% NDF starter (p = 0.001) in the immediate post-weaning and isovaleric acid which was greater in early-weaned calves (p = 0.044) at the end of experiment. Serum metabolites were largely affected (p < 0.05) by starter NDF levels and weaning time in the pre-weaning phase only. Alpha diversity of rumen microbes was greater and chaotic in 14% NDF starter group (early- and late-weaned) in the pre-weaning phase which converged in the immediate post-weaning phase and diverged on starter NDF basis at the end of experiment. Microbial ecology at phylum and genus levels composition were greatly driven by starter NDF levels in the pre-weaning phase, by weaning time in the immediate post-weaning phase, and two distinct bifurcated microbial ecologies based on starter NDF levels appeared at the end of experiment. In conclusion, the comparable growth with distinct microbial diversity but largely in favor of 24% NDF starter suggests that calves can be subjected to early weaning with 24% starter NDF levels for smooth transition from liquid to solid feed in Holstein calves. Full article

This study evaluated the effects of Nypa fruticans fruit pellet supplementation combined with different CP levels on rumen fermentation characteristics and CH₄ production using an in vitro gas production technique. A 3 × 4 factorial arrangement was used, consisting of three CP levels (12, 14, and 16%) and four levels of Nypa fruticans fruit pellet supplementation (0, 0.5, 1.0, and 1.5% of substrate dry matter), with incubation run included as a random effect in the statistical model. Rumen fluid from Thai native beef cattle was incubated under anaerobic conditions. Gas production kinetics, ruminal pH, ammonia–nitrogen (NH₃–N), protozoal populations, digestibility, volatile fatty acids (VFA), and CH₄ production were determined. Significant interactions between CP level and Nypa fruticans fruit pellet supplementation were observed for gas production kinetics. Ruminal pH was influenced by CP level at 24 h, while NH₃–N increased with higher CP levels but decreased with increasing supplementation. Protozoal populations were reduced by Nypa fruticans fruit pellets. Methane production was affected by CP level, Nypa fruticans fruit pellet supplementation, and their interaction. A clearer reduction was observed at 24 h, particularly at higher supplementation levels. At 24 h of incubation, total VFA, propionate, and butyrate concentrations increased with supplementation, whereas no clear effects were observed at 12 h. In vitro dry matter digestibility was affected at 24 h (p < 0.05), but no effect was observed at 48 h, while organic matter digestibility remained unchanged. In conclusion, Nypa fruticans fruit pellets, in combination with CP level, modified rumen fermentation patterns and were associated with lower CH₄ production under in vitro conditions, without negatively affecting digestibility. These findings suggest potential for further in vivo evaluation. Full article

This study aims to evaluate the blanching process of wasted Undaria pinnatifida as a ruminant feed source by assessing chemical compositions, in vitro nutrient digestibility, rumen fermentation characteristics, greenhouse gas emissions, and rumen microbes. The blanching process was conducted at different temperatures (15 vs. 80 vs. 90 °C) and times (2 vs. 4 min) to assess the chemical and mineral contents. Supplementation levels of U. pinnatifida (0 vs. 0.5 vs. 1 vs. 2%) were observed with the blanching process (non-blanching (NBL) vs. blanching (LOS)). Increasing blanching temperature and time decreased (p < 0.05) dry matter, crude ash, and the mineral contents, including sodium, phosphorus, and arsenic. Moreover, LOS treatment increased (p < 0.01) in vitro dry matter and neutral detergent fiber digestibility, ruminal pH, and the acetate-to-propionate ratio, but reduced (p < 0.01) CH₄ (mL/g NDFD). Additionally, 2% of LOS treatment reduced the abundance of protozoa, fungi, fibrolytic microbes, methanogenic archaea, Methanobrevibacter ruminantium, Methanosarcina barkeri, and Methanosphaera stadtmanae (p < 0.01). Therefore, blanching at 80 °C for 2 min improved the nutritional profile by reducing antinutritional minerals. Subsequent in vitro fermentation suggested that supplementing the diet with 0.5–1% of LOS improved digestibility and altered fermentation, potentially reducing methane yield (per NDFD). Full article

This study investigated the interactive effects of corn silage and ramie silage on in vitro rumen fermentation characteristics, aiming to provide a scientific basis and empirical evidence for the rational incorporation of ramie into ruminant diets. Four binary substrate mixtures were formulated based on dry matter (DM) mass ratios of corn silage to ramie silage: 100:0 (CON), 60:40 (R40), 20:80 (R80), and 0:100 (R100). Rumen fluid was collected from three adult Liuyang black goats surgically fitted with permanent rumen cannulas, and a standardized 48 h in vitro batch culture assay was conducted. Results demonstrated that increasing the proportion of ramie silage significantly decreased (p < 0.05) the DM degradation rate, neutral detergent fiber (NDF) degradation rate, acid detergent fiber (ADF) degradation rate, and total gas production per gram of substrate DM. Specifically, CON and R40 exhibited significantly higher values for all four parameters than R80 and R100 (p < 0.05). Methane production was significantly reduced in all ramie-containing treatments relative to CON (p < 0.05), whereas hydrogen production increased progressively with ramie inclusion level, with CON yielding significantly less H₂ than both R80 and R100 (p < 0.05). Regarding fermentation parameters, increasing ramie proportion elevated (p < 0.05) both fermentation fluid pH and the acetate-to-propionate ratio, while total volatile fatty acid (TVFA) concentration declined linearly (p < 0.05). TVFA concentrations did not differ significantly between CON and R40, yet both were significantly greater than those in R80 and R100 (p < 0.05). Collectively, these findings indicate that ramie silage is a nutritionally valuable forage with potential as a high-quality partial replacement for conventional silages in ruminant feeding systems; however, its inclusion in corn–ramie mixed silages should not exceed 40% (on a DM basis) to maintain optimal fermentative efficiency and nutrient degradability. Full article

Rumen microbial fermentation plays a central role in nutrient utilization and milk production in dairy cows. This study evaluated the effects of supplementation with exogenous fibrolytic enzymes, alone or in combination with live yeast on rumen microbiota, fermentation characteristics, nitrogen-related metabolites, and production performance during the transition from outdoor grazing to indoor housing. Thirty Lithuanian Red dairy cows were assigned to control (CTR), enzyme (E), or enzyme plus yeast (YE) treatments across outdoor (OD) and transit (T) periods, while nine cows (three per group) were selected for rumen and microbiota analysis. Rumen bacterial communities were characterized using 16S rRNA gene sequencing, and functional parameters were evaluated using linear mixed-effects models. Supplementation resulted in selective changes in several bacterial genera, including Blautia spp., WPS-2, Ruminococcus spp., Erysipelotrichaceae UCG-009, Sharpea spp., uncultured Bacteroidales, and Prevotellaceae UCG-003, and was associated with alterations in fermentation patterns, particularly propionate concentration, and in nitrogen metabolism, including putrescine dynamics. The transition period significantly influenced microbial diversity and total bacterial abundance across treatments. Cows in the YE group maintained higher milk yield during the transition period. Overall, dietary supplementation modulated specific rumen metabolic responses and contributed to production stability without causing large-scale changes in overall microbial structure. Full article

In ruminant nutrition, the lignocellulosic complex is a primary constraint limiting the utilization of dietary fiber. The objective of this study was to evaluate the effects of inoculating lignin-degrading bacteria (LDB) isolated from the ruminant rectum on in vitro rumen fermentation characteristics. Rectal fecal samples were collected from healthy beef cattle, dairy cattle, buffaloes, and goats (n = 4 per species) using the grab sampling technique. Twenty-eight bacterial colonies were isolated through enrichment and screening on media containing sodium lignosulfonate. Lignin degradation efficiency was assessed spectrophotometrically, while laccase activity was determined using a 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) oxidation assay. Seven isolates exhibiting ligninolytic activity (1.4–5.6% degradation efficiency) were selected to evaluate their effects on in vitro rumen fermentation using a completely randomized design with four replicates. LDB treatments were standardized to a concentration of 2.4 × 10⁵ colony-forming units/mL of rumen fluid medium, while the control received an equal volume of a 0.85% sterile NaCl solution. A rice straw-based total mixed ration served as the substrate, with rumen fluid collected from beef cattle. All treatments were incubated for 48 h. Notably, isolate BC3 consistently enhanced in vitro dry matter digestibility (23.1%), total gas production (18.6%), and total volatile fatty acid concentrations (13.2%) relative to the control and other LDB isolates (p < 0.01). All seven LDB isolates were identified as Gram-negative, rod-shaped, facultative anaerobic bacteria that exhibit catalase activity and tolerate moderately acidic conditions. Phylogenetic tree analysis based on 16S rRNA gene sequencing identified isolate BC3 as being closely related to Escherichia coli strains. These findings demonstrate that the ruminant hindgut is a promising source of LDB with the functional potential to enhance feed digestibility and fermentation end-products in the rumen. Future research should prioritize in vivo trials to evaluate the safety and efficacy of LDB as a direct-fed microbial, specifically focusing on its impact on animal performance and health. Full article

The present study was conducted to demonstrate the effects of Codium fragile extract on methane production using in vitro and in vivo experiments. An in vitro batch experiment was conducted to evaluate different inclusion levels of Codium fragile extract (0, 0.25, and 0.5% of substrate dry matter). Methane production significantly decreased in the 0.5% treatment (p < 0.05), whereas dry matter digestibility and total volatile fatty acid concentration were not significantly affected (p > 0.05). Based on the in vitro results, an in vivo feeding experiment was conducted using a 0.5% inclusion level of Codium fragile extract on Hanwoo steers. Methane emissions were significantly decreased by approximately 10% in steers fed Codium fragile extract (p < 0.05). In contrast, rumen fermentation characteristics, feed intake, average daily gain, and blood parameters were not significantly different between the treatments (p > 0.05). These results demonstrate that a dietary additive with 0.5% Codium fragile extract effectively reduced methane emissions without negatively affecting rumen fermentation and growth performance in Hanwoo steers. Full article

The sensory quality and flavor of lamb meat, critical to market competitiveness, are influenced by rumen microbial fermentation and dietary management strategies. Coated betaine (CBet), a rumen-protected methyl donor, exerts systemic nutritional regulation in ruminants. This study explored the effects of CBet supplementation on lamb meat quality using 18 Dorset ♂ × Hu sheep ♀ F1 crossbred lambs, randomly assigned to either a control group (basal diet) or a 0.20% CBet-supplemented diet for 60 days (n = 9 per group). The results demonstrated that CBet significantly increased ruminal concentrations of total volatile fatty acids (TVFAs), acetic acid, propionic acid, and butyric acid (p < 0.05). Additionally, CBet supplementation enhanced muscle redness (a*), crude fat, crude ash, heptadecanoic acid (C17:0), and tricosanoic acid (C23:0) (p < 0.05) while decreasing shear force and the concentration of cis-13,16-docosadienoic acid (C22:2) (p < 0.05). Furthermore, CBet elevated characteristic flavor compounds (e.g., nonanal) and their relative odor activity values (ROAVs), and decreased undesirable odors (e.g., dodecanal) (p < 0.05). As illustrated in the graphical abstract, these improvements were mediated through regulatory effects of CBet on rumen microbiota composition, muscle fatty acids, amino acids, and volatile flavor compounds. Specifically, CBet significantly increased the relative abundances of Firmicutes, Proteobacteria, Prevotella, and Bifidobacterium in the rumen (p < 0.05) and altered the Firmicutes/Bacteroidota ratio. In conclusion, dietary supplementation with 0.20% CBet effectively enhances lamb meat quality and flavor, effects closely associated with changes in the abundance of key ruminal microbial taxa. Full article

This study investigated the effects of isoacid supplementation on in vitro rumen fermentation characteristics, nutrient degradability, and bacterial community diversity in yaks using corn silage–highland barley straw-based substrates. An in vitro fermentation experiment was conducted with a substrate consisting of 80% whole-plant corn silage and 20% highland barley straw. Treatments included a control (without isoacids) and four isoacid supplemental levels (0.1%, 0.2%, 0.3%, and 0.4% of substrate dry matter, DM), each with six replicates. A 72 h in vitro gas production experiment was performed to measure cumulative gas production, fermentation parameters, nutrient degradability, and bacterial community diversity. Cumulative gas production increased by 12.96% with 0.2% isoacid supplementation compared to the control (p < 0.05). The contents of microbial protein (MCP), acetate, propionate, and total volatile fatty acids (TVFA) exhibited quadratic responses to the increasing isoacid dosage (p < 0.05). Specifically, MCP content reached a maximum of 0.76 mg/mL with 0.2% isoacids, representing a 31.03% increase compared to the control (p < 0.05). TVFA content was highest (146.85 mmoL/L) at 0.2% isoacid supplementation, with a 16.40% increase compared to the control (p < 0.05). Acetate content increased by 17.99% (p < 0.05), while propionate tended to increase with 0.2% isoacid supplementation (p = 0.08). Supplementation with 0.2% and 0.4% isoacids did not alter the bacterial composition and diversity (p > 0.05). However, at the genus level, g_Ruminococcus, g__Elusimicrobium, g_norank_f_Atopobiaceae, g_norank_o_Coriobacteriales, and g_Romboutsia were identified as differential biomarkers showing significant responses to isoacid supplementation (p < 0.05). Mantel-test analysis revealed positive correlation between g_Ruminococcus abundance and NH₃-N content (r < 0.4, p < 0.05); g_Romboutsia abundance and acetate content (r < 0.40, p < 0.05); g_Defluviitaleaceae_UCG-011 abundance and both NH₃-N content and the pH of rumen fluid (r < 0.40, p < 0.05); g_norank_o_Coriobacteriales abundance and rumen pH (r < 0.40, p < 0.01). Supplementation with 0.2% isoacids to corn silage–barley straw substrates improved in vitro rumen fermentation characteristics in yaks, which was associated with altered abundances of key bacterial genera including g_Ruminococcus, g__Elusimicrobium, g_norank_f_Atopobiaceae, g_norank_o_Coriobacteriales. Full article