Search Results (327)

Global warming has become a significant public health concern, with intensive livestock farming as a major contributor. To mitigate greenhouse gas emissions, strategies such as manipulating the ruminal environment with dietary additives are essential. This study evaluated Moringa oleifera, a globally widespread tree with antioxidant, multivitamin, protein-rich, and anti-inflammatory properties, as a feed additive. Rumen fluid was collected from three Pelibuey sheep, homogenized, and subjected to an in vitro fermentation study for 48 h with three alfalfa/moringa ratio treatments: T0 Control (100:0), T1 Low (85:15), and T2 High (70:30). Total RNA was extracted, followed by high-definition sequencing of the metatranscriptome. The sequencing yielded approximately 456 million sequences. A total of 117 phyla were identified and approximately 1300 genera were mapped. Predominant phylum differed by treatment: T0, Firmicutes; T1, Proteobacteria; and T2 with Synergistetes, at least one sample per treatment. Archaea were nearly absent in T1, which explains a statistically significant decrease in methane production. In the Gene Set Enrichment Analysis (GSEA), it was observed that one of the metabolic pathways with a statistically significant difference (p-value < 0.05) was that of methane, specifically in the low moringa treatment (T1) compared to the control (T0). From the functional analysis, differentially expressed enzymes were identified, some of which are involved in the methane metabolic pathway, such as formate dehydrogenase (EC 1.17.1.9) and glycine hydroxymethyltransferase (EC 2.1.2.1), which are intermediates in methane formation. These results suggest that 15% Moringa oleifera supplementation alters ruminal microbiota, reduces archaeal activity, and suppresses methane-related pathways. These findings provide molecular evidence supporting the potential of M. oleifera as a methane mitigation strategy in ruminant nutrition. Full article

Okara, a protein-rich byproduct of soymilk production, is highly perishable because of its high moisture content. This study evaluated the preservation and nutritional value of okara fermented by lactic acid bacteria for use in dairy cattle diets. Fermentation effectively reduced pH within 2 weeks and maintained quality for up to 6 weeks. However, aerobic exposure increased the concentration of ammonia, indicating a decline in stability. In vitro assessments revealed no significant differences in in vitro true dry matter digestibility, in vitro neutral detergent fiber digestibility, or gas production between fermented and fresh okara, although fermented okara had a higher concentration of ammonia nitrogen. In situ analysis revealed slightly lower dry matter effective degradability (ED) in fermented okara, but similar rumen-degradable and undegradable protein fractions. When fermented okara was used to replace soybean meal in total mixed rations, 25–50% inclusion-maintained digestibility and fermentation characteristics, with 25% replacement yielding the highest ED at a low ruminal passage rate (0.02 h⁻¹). Taken together, these results suggest that fermented okara can be strategically incorporated into dairy rations as a sustainable protein alternative, supporting both rumen function and bypass protein supply. Full article

Prickly pear by-products contain dietary fibre and bioactive components like polyphenols and flavonols, which can reduce total gas and methane emissions. To this end, an in vitro trial was carried out in duplicate utilizing three diets containing hay, concentrate, and two prickly pear by-products obtained after grinding the fruit peel and pastazzo (pulp + peel + seeds), which were ensiled with the addition of 12% wheat bran (raw weight). Based on the ingredient intake recorded in the in vivo study for 12 lactating ewes fed the three diets, an in vitro rumen fermentation study with the innovative Gas Endeavour system (GES) was performed, and the Gage R&R statistical method was used to evaluate the accuracy of the total gas and methane production detected by the GES device. Fermented liquor samples for each diet were used to calculate the disappearance of organic matter and neutral detergent fibre. Shotgun metagenome sequencing analysis was used to evaluate the effect of diet on the rumen fluid microbiota, and it was found that the parameters of repeatability and reproducibility of the total gas and the methane produced after 24 h were satisfactory. Prickly pear by-products display high fermentability for the peel and low fermentability for pastazzo silage, which generates lower total gas and methane emissions. This diminished methane gas production is not correlated with the relative abundance of methanogens. The different chemical and nutritional composition of the three diets altered the rumen bacteria, albeit only slightly, with particular reference to the Succinivibrio and Selenomonas genera. In conclusion, prickly pear peel silage displayed acceptable fermentation traits, which could support its utilization in sheep diets. Full article

The present experiment aimed to investigate the effects of harvest stages on the fermentation quality and nutritional value of sorghum stalk silage. Sorghum stalks were harvested at the three stages (milk, dough, and ripe), chopped, and ensiled for 60 d. Each treatment had five replicates, and the silages were evaluated for fermentation quality, nutritional composition, in vitro rumen fermentation characteristics, and bacterial community profiles. The results showed that the crude protein, neutral detergent fiber, and acid detergent fiber contents decreased significantly with harvest maturity (p < 0.05). Consequently, silage from the ripe stage possessed the highest dry matter, relative feed value, and total digestible nutrients (p < 0.05). In vitro rumen fermentation indicated that the ripe stage silage exhibited the greatest gas production, and the lowest concentrations of ruminal ammonia–nitrogen (p < 0.05). Microbial analysis revealed a shift from dominant epiphytic Proteobacteria to fermentative Firmicutes post-ensiling, with the ripe stage community co-dominated by Lactobacillus and Leuconostoc, in contrast to the milk stage’s enrichment with Klebsiella. In conclusion, harvesting sorghum at the ripe stage is the optimal strategy as it establishes a beneficial microbial community, resulting in silage with superior nutritional value and rumen fermentation efficiency. Full article

Whole-plant corn silage is a critical feedstuff in global ruminant production, and its nutrient composition is closely tied to harvest timing. As starch acts as the primary energy source in silage-based diets, investigating changes in starch degradation rate provides a theoretical basis for optimizing the efficient utilization of whole-plant corn and its silage in ruminant production. In this study, whole-plant corn (harvested from the milk stage to full ripening stage) and its corresponding silage were used as experimental materials. An in vitro simulated rumen fermentation system was employed to determine the contents of starch, prolamin, amylose, and amylopectin in the samples. The results showed that with delayed harvest time, starch content in both whole-plant corn and its silage increased significantly; prolamin and amylose contents first decreased, then increased; amylopectin content first rose significantly before decreasing; and both starch disappearance rate and speed exhibited a trend of first increasing, then decreasing. After silage fermentation, the silage had significant increases in starch, amylose, and amylopectin contents, and starch disappearance rate; prolamin content decreased; and starch disappearance speed increased extremely significantly. This study indicates that whole-plant corn harvest time and silage fermentation regulate the ruminal starch degradation pattern by altering starch structure, prolamin content, and the proportion of rapidly degradable starch. Full article

Methane (CH₄), originating from ruminants, is a major source of greenhouse gas emissions in the agriculture industry. This study aimed to determine the potential of red seaweed Gracilaria lemaneiformis (G. lemaneiformis) as an anti-methanogenic feed additive for cattle. Three supplementation levels of seaweed (2%, 5%, and 10% of dry matter) were evaluated for their effects on gas production and rumen fermentation characteristics during 48 h in vitro fermentation. The results revealed a significant decrease in total gas production (TGP), CO₂, CH₄, ammonia nitrogen (NH₃-N), and volatile fatty acid (VFA) concentrations, with no differences in pH or dry matter disappearance (DMD). Notably, compared with the control group without seaweed, supplementation with 2% G. lemaneiformis effectively reduces CH₄ emissions by 27.5% (p < 0.05). Supplementation with 2% G. lemaneiformis decreased the abundance of methanogens g_norank_f_Methanomethylophilaceae, responsible for CH₄ generation, and increased the populations of bacteria (Kandleria and Succinivibrio) that compete with methanogens for substrates. Furthermore, upregulating the levels of 13(S)-HOTrE and 9(S)-HOTrE (polyunsaturated fatty acids) could inhibit methanogenic activity. Additionally, lower VFA concentrations will provide less raw materials for methane synthesis, thus further inhibiting methanogenesis. In summary, G. lemaneiformis, as a red seaweed with important economic value, can not only be applied to enhance marine carbon sinks but can also serve as a promising candidate for mitigating biomethane emissions in cattle. Full article

This study evaluated the effects of replacing SBM with CWYWEP on in vitro rumen fermentation, nutrient degradability, and gas production kinetics. Citric waste was co-fermented with yeast waste and a multi-enzyme complex for 14 days, then sun-dried and pelleted. The final CWYWEP product contained 50.4% crude protein (DM basis). A completely randomized design tested seven diets in which SBM was replaced by CWYWEP or non-enzymatic citric waste–yeast waste pellets (CWYWP) at 0%, 33%, 66%, or 100% inclusion. Replacing SBM with CWYWEP significantly increased cumulative gas production at 96 h, with the 100% CWYWEP group achieving 93.7 mL/0.5 g DM—a 14% increase over the control (p < 0.01). Microbial lag time was reduced to 0.17 h vs. 0.28 h in the control (p < 0.05), suggesting faster microbial colonization. The highest in vitro DM degradability (IVDMD) at 48 h was observed in the 100% CWYWEP group (64.5%), outperforming both the SBM control and all CWYWP treatments (p < 0.01). Notably, CWYWEP increased total volatile fatty acids by 5% at 4 h and propionate by 9% at 2 h, while reducing methane production by 5% (p < 0.05). Other parameters, including pH, ammonia nitrogen, organic matter digestibility, and protozoal counts, were unaffected (p > 0.05). In contrast, CWYWP without enzymes showed minimal improvement. These findings indicate that CWYWEP is a promising high-protein alternative to SBM, enhancing fermentation efficiency and reducing methane under in vitro conditions. Further in vivo studies are warranted to validate these effects. Full article

Enhancing forage protein is key to sustainable ruminant nutrition. The nutritive value of Hedysarum coronarium L. was investigated by studying different preservation systems (fresh vs. dehydrated pellet vs. hay) (Exp. 1) and morphological fractions (flowers vs. leaves vs. stems) (Exp. 2). For the fresh and pelleted systems, two cuts were used. Proximately, total polyphenols and condensed tannins were detected. In vitro fermentation characteristics were studied by incubating samples with buffered sheep rumen fluid, estimating methane production by volatile fatty acids. Fresh and pelleted sulla were more nutritionally advantageous than hay, in terms of metabolizable energy and protein-to-fiber ratio. Pelleting at the beginning of flowering proved to be a suitable forage for sheep feeding due to protein (15.1% DM), metabolizable energy (9.64 MJ/kg DM), structural carbohydrates (39.5% DM), and total polyphenols (13.5 GAE g/kg DM) content. This cut showed an in vitro fermentation rate (9.86 mL/h), organic matter degradability (55.7%), and volatile fatty acids (87.3 mmoL/g) that were higher (p < 0.05) than hay. Flowers and leaves showed higher (p < 0.05) levels of secondary metabolites than stems as well as lower methane production. These results suggest the potential influence of these compounds in reducing rumen emissions. Dehydration and pelleting resulted in an effective preservation method for maintaining nutrients in sulla forage. Full article

While the Ankom RF system facilitates efficient high-throughput in vitro fermentation studies, its high cost and limited flexibility constrain its broader applicability. To address these limitations, we developed and validated a low-cost, modular gas monitoring system (FerME), assembled from commercially available components. To evaluate its performance and reproducibility relative to the Ankom RF system (Ankom Technology, Macedon, NY, USA), in vitro rumen fermentation experiments were conducted under strictly controlled and identical conditions. Whole rumen contents were collected approximately 2 h post-feeding from individual mid- or late-lactation dairy cows and immediately transported to the laboratory. Each fermenter received 50 mL of processed rumen fluid, 100 mL of anaerobically prepared artificial saliva buffer, and 1.2 g of the donor cow’s diet. Bottles were sealed with the respective system’s pressure sensors, flushed with CO₂, and incubated in a 50 L water bath maintained at 39 °C. FerME (New Taipei City, Taiwan) and Ankom RF fermenters were placed side-by-side to ensure uniform thermal conditions. To assess the effect of filter bag use, an additional trial employed Ankom F57 filter bags (Ankom Technology, Macedon, NY, USA; 25 μm pore size). Trial 1 revealed no significant differences in cumulative gas production, volatile fatty acids (VFAs), NH₃-N, or pH between systems (p > 0.05). However, the use of filter bags reduced gas output and increased propionate concentrations (p < 0.05). Trial 2, which employed filter bags in both systems, confirmed comparable results, with the FerME system demonstrating improved precision (CV: 4.8% vs. 13.2%). Gas composition (CH₄ + CO₂: 76–82%) and fermentation parameters remained consistent across systems (p > 0.05). Importantly, with 12 pressure sensors, the total cost of FerME was about half that of the Ankom RF system. Collectively, these findings demonstrate that FerME is a reliable, low-cost alternative for real-time rumen fermentation monitoring and could be suitable for studies in animal nutrition, methane mitigation, and related applications. Full article

This study evaluated the effects of liquid oregano oil, chitosan nanoparticles with encapsulated liquid oregano oil, and a negative control of empty chitosan nanoparticles on in vitro ruminal fermentation. Three Boer goats were used as ruminal fluid donors, fed with a formulated ration for 21 d for inoculum adaptation. Treatments tested on in vitro assays were diet without oregano oil or nanoparticles (CON); diet with 100 ppm of oregano oil in nanoparticles (100N); diet with 300 ppm of liquid oregano oil (300L); diet with 300 ppm of oregano oil in nanoparticles (300N); and diet with 300 ppm of empty nanoparticles (300CHN). The variables studied were in vitro dry matter digestibility (ivDMD), in vitro neutral detergent fiber digestibility (ivNDFD_om), total gas production (TGP), ammonia nitrogen concentration (NH₃), and pH. The experimental design was a randomized complete block design. Linear and quadratic regressions were used to identify dependence and inflection points. The ivDMD increased at 12, 36, and 48 h, with 300N and with 300L exhibiting increased ivNDFD_om at 36 h. Ruminal pH was highest (p < 0.05) with 300CHN at 36 h. For first-order regression analysis of TGP, coefficients (β) were highest (p < 0.05) for 300N. In conclusion, 300N increased ruminal fermentation in vitro, as reflected by increases in ivDMD, ivNDFD_om, and TGP. Full article

Feed supplementation strategies are essential for optimizing cattle productivity, and the incorporation of non-conventional feed resources may reduce both production costs and environmental impact. This study evaluated the effects of pelletized protein concentrates (including Acacia farnesiana, A. schaffneri, and Agave duranguensis bagasse) on rumen fermentation parameters, microbial communities, and gas emissions. Fistulated bullocks received the concentrate daily, and ruminal contents were collected and filtered before and after supplementation to assess in vitro gas and methane production, pH, and microbial composition using high-throughput sequencing of 16S rRNA and mcrA amplicons. In addition, in situ degradability was evaluated during and after the supplementation period. Supplementation led to a significant (p < 0.05) reduction in degradability parameters and methane production, along with a marked decrease in the abundance of Methanobrevibacter and an increase in succinate-producing taxa. These effects were attributed to the enhanced levels of non-fiber carbohydrates, hemicellulose, crude protein, and the presence of bioactive secondary metabolites and methanol. Rumen microbiota composition was consistent with previously described core communities, and mcrA-based sequencing proved to be a valuable tool for targeted methanogen detection. Overall, the inclusion of non-conventional ingredients in protein concentrates may improve ruminal fermentation efficiency and contribute to methane mitigation in ruminants, although further in vivo trials on a larger scale are recommended. Full article

The purpose of this study was to explore the effects of co-ensiling Caragana korshinskii with different proportions of oat grass on silage fermentation quality, chemical composition, in situ rumen degradability and in vitro rumen fermentation characteristics. C. korshinskii and oat grass were mixed at different ratios of 100:00, 90:1, 80:2, 70:30, 60:40 and 50:50. Each ratio of mixture was ensiled for 7, 14, 30, 45 and 60 days at room temperature (25 °C), with 30 bags per ratio, for a total of 180 bags. We further investigated the dynamic profiles of the bacterial community during ensiling and in vitro rumen fermentation. The results showed that co-ensiling C. korshinskii and oat grass decreased the pH values and increased the content of lactic acid and acetic acid compared with ensiling C. korshinskii alone. C. korshinskii ensiled with oat grass at a ratio of 70:30 (70% C. korshinskii) showed the best fermentation quality, which was related to higher relative abundance of Lactobacillus and Weissella. The silage with the ratio of 70:30 (70% C. korshinskii) showed higher dry matter digestibility and the more production of gas and total volatile fatty acids, compared with fresh C. korshinskii. In conclusion, C. korshinskii co-ensiled with oat grass at a ratio of 70:30 could enhance the fermentation quality and digestibility of C. korshinskii. Full article

This study aimed to evaluate the effect of including silage from devilfish waste (SF-Hypostomus plecostomus) and probiotics (PB-Pediococcus acidilactici BX-B122 and Bacillus coagulans BX-B118) in ruminants on greenhouse gas production. The diets evaluated contained 0, 8, 14 and 20% of silage made from SF and the addition of PB at a dose of 0.2 mL/g of diet, using steers and sheep (rams) as rumen inoculum donors in a completely randomized statistical design with a 2 × 4 × 2 factorial arrangement. Asymptotic gas production (GP) was influenced (p < 0.01) by the interactions between rumen liquor (RL), SF, and PB. The inclusion of SF and PB resulted in a higher (p < 0.01) GP rate in sheep; however, the values were reduced with increasing levels of SF. Asymptotic CH₄ in the rumen fluid of steers decreased with an increasing SF percentage up to 14%. Probiotics had different effects on the rumen fluid of sheep and steers. In steers, probiotics substantially reduced (p < 0.01) CH₄ synthesis while supplementation increased it in sheep rumen fluid. Similarly, diets with probiotics had higher CO formation (p < 0.05) in sheep and steer liquor. Similarly, CO decreased (p < 0.05) with increasing levels of SF. In the rumen fluid of sheep and steers, the probiotics were found to reduce H₂S, while there was an SF-dose-dependent decrease in H₂S concentration. The ruminal pH and dry matter digestibility of sheep were higher than in steers. It can be concluded that increasing SF levels generally reduced the total gas and CH₄ production, with probiotics further enhancing this reduction, especially in CH₄ per unit of gas. Full article

Ruminant livestock contribute significantly to methane emissions, necessitating sustainable mitigation strategies. Essential oils (EOs) show promise for modulating ruminal fermentation, but their synergistic effects remain underexplored. Two 24 h in vitro experiments evaluated the synergistic effects of EO blends on rumen microbial fermentation. Exp. 1 screened five oils using two triad combinations. Triad 1 tested 10 combinations of thyme (THY), peppermint (PPM), and cinnamon leaf (CIN) oils. Triad 2 tested 10 combinations of anise (ANI), clove leaf (CLO), and peppermint (PPM) oils. Each blend was tested at 400 mg/L, using batch culture methods measuring: pH, ammonia-N (NH₃-N), and volatile fatty acids (VFAs). The two most effective blends, designated as T1 and T2, were selected for Exp. 2 to assess total gas and methane (CH₄) production using pressure transducer methods. All treatments were incubated in a rumen fluid–buffer mix with a 50:50 forage-to-concentrate substrate (pH 6.6). In Exp. 1, data were analyzed according to the Simplex Centroid Design using R-Studio. In Exp. 2, an analysis was conducted using the MIXED procedure in SAS. Mean comparisons were assessed through Tukey’s test. The results from Exp. 1 identified CIN+PPM (80:20) and ANI+CLO (80:20) as optimal combinations, both increasing total VFAs while reducing acetate/propionate ratios and NH₃-N concentrations. In Exp. 2, both combinations significantly reduced total gas and CH₄ productions compared to the control, with CIN+PPM achieving the greatest methane reduction (similar to monensin, the positive control). Specific essential oil combinations demonstrated synergistic effects in modulating rumen fermentation and reducing methane emissions, offering potential for sustainable livestock production. Further in vivo validation is required to optimize dosing and assess long-term effects on animal performance. Full article

The aims of the current study were to characterize the natural compounds of Salsola tragus via GC-MS and determine its effects as a forage source on lamb growth, in vitro rumen fermentation kinetics, carcass quality, cellular damage, and metabolomic meat interaction. Twenty-one Rambouillet lambs were randomly assigned to one of three experimental diets (seven lambs per treatment): (1) a control diet (W/o-Salsola) containing 300 g/kg dry matter (DM) of sorghum stover; (2) a diet with a medium inclusion of Salsola tragus (15-Salsola), which contained 150 g/kg DM of Salsola tragus and 150 g/kg DM of sorghum stover; and (3) a diet comprising 300 g/kg of Salsola tragus (30-Salsola). The results showed there were no differences (p > 0.05) in lamb growth performance during the fattening phase. The in vitro gas analysis demonstrated that the 30-Salsola treatment increased lag time h⁻¹ (p < 0.05) and reduced gas production (p = 0.03). The metabolomic analysis findings suggest that the treatments that included Salsola tragus significantly positively affect the metabolomic composition of meat (p < 0.05). The use of 15-Salsola as a source of forage is promising for feeding lambs during the fattening phase. Full article