Search Results (283)

High-molecular-weight polycyclic aromatic hydrocarbons (PAHs), such as pyrene, are persistent environmental pollutants that threaten soil health and agricultural productivity due to their resistance to degradation. This study evaluated the efficacy of domesticated bacteria isolated from contaminated farmland soil and activated sludge, used alone and in combination with corn (Zea mays L.), to remove pyrene from soil, enhance plant growth, improve tolerance, and ensure crop safety. Six bacterial strains were isolated: three from polluted farmland soil (WB1, WB2, and WF2) and three from activated sludge (WNB, WNC, and WH2). High-throughput 16S rRNA amplicon sequencing profiled bacterial communities after 30 days of treatment. Analytical tools, including LEfSe, random forest, and ZiPi analyses, identified biomarkers and core bacteria associated with pyrene degradation, assessing their correlations with plant growth, tolerance, and pyrene accumulation in corn straw. Bacteria from activated sludge (WNB, WNC, and WH2) outperformed farmland soil-derived strains and the inoculant strain ETN19, with WH2 and WNC achieving 65.06% and 87.69% pyrene degradation by days 15 and 30, respectively. The corn–bacteria consortium achieved up to 97% degradation. Activated sewage sludge (ASS)-derived bacteria were more effective at degrading pyrene and enhancing microbial activity, while soil-derived bacteria better promoted plant growth and reduced pyrene accumulation in straw. Microbial communities, dominated by Proteobacteria, exhibited high species richness and resilience, contributing to xenobiotic degradation. The corn-domesticated bacteria consortia effectively degraded pyrene, promoted plant growth, and minimized pollutant accumulation in crops. This remediation technology offers a promising strategy for rapid and sustainable bioremediation of agricultural soils contaminated with organic compounds such as PAHs or other complex pollutants, while promoting the development of efficient bacterial communities that enhance crop growth. Full article

Endophytic fungi represent key microbial symbionts that colonize internal plant tissues without causing apparent disease, playing vital roles in host growth, stress resistance, and biosynthesis of bioactive compounds. Carpesium lipskyi C. Winkl., a medicinal plant endemic to the Gaoligong Mountains in Yunnan, remains largely unexplored regarding its endophytic fungal composition. In this study, a total of 737 amplicon sequence variants (ASVs) were identified through high-throughput sequencing, spanning 9 phyla, 36 classes, 67 orders, 137 families, 206 genera, and 277 species. The dominant phyla were Ascomycota, Basidiomycota, and Glomeromycota. Alpha diversity in stems and leaves followed a unimodal distribution along the elevational gradient, in contrast to root endophytic communities, which showed no significant correlation with altitude. Peak diversity occurred at 2734 m, indicating a non-linear altitude-diversity relationship. Altitude, along with stable precipitation and temperature (2600–3210 m), significantly influenced fungal diversity. Medicinal fungi such as Cladosporium sp., Meyerozyma guilliermondii, Phialocephala fortinii, and Rhodotorula mucilaginosa were found in either roots or stems. This is the first comprehensive assessment of endophytic fungi in C. lipskyi from this region, providing a foundation for future ecological and pharmacological studies. Full article

Soil microbial communities are important for nutrient cycling regulation in forest ecosystems. However, limited knowledge exists regarding the characteristics of these microbial communities in Masson pine (Pinus massoniana Lamb.) plantations of different stand ages. In this study, four planted Masson pine stands (8-year-old, 12-year-old, 22-year-old, and 38-year-old stands) and one natural broadleaved forest stand (as a control) with three replications, were selected in the Laoshan Forest Farm, Qiandao Lake Town, Zhejiang Province, China. Soil physicochemical properties were measured and their effects on soil microbial communities were studied. Amplicon-based high-throughput sequencing was employed to process raw sequence data for soil microbes. It is worth noting that significant differences (p < 0.05) in soil bacterial genera were observed among different stand age groups. Total nitrogen (TN), total phosphorus (TP), total potassium (TK), available potassium (AK), soil organic carbon (SOC), and soil bulk density (BD) were identified as the primary factors influencing bacterial community distribution (p < 0.05). Available nitrogen (AN), SOC, TN, and TK showed significant correlations with soil fungal communities (p < 0.05). These findings underscore the crucial role of soil physicochemical properties in shaping soil microbial community composition in Masson pine plantations. Full article

Endophytic bacteria play an important role in the growth, stress tolerance, and metabolic function of salt-tolerant peanuts, yet their community assembly across different saline–alkali soils and plant organs remains poorly characterized. In this study, the V3–V4 variable region of the endophytic bacteria 16S rRNA gene in three organs (roots, leaves, and pods) of high-oleic-acid peanut variety Huayu9118 from three saline–alkali locations (Xinjiang, Jilin, and Shandong, China) was analyzed by high-throughput sequencing. A total of 1,360,313 effective sequences yielded 19,449 amplicon sequence variants (ASVs), with Proteobacteria (45.86–84.62%), Bacteroidota (6.52–13.90%), and Actinobacteriota (3.97–10.87%) dominating all samples. Niche strongly influenced microbial diversity: the roots exhibited the highest level of richness (Chao 1/ACE indices), while the leaves showed the greatest diversity (Shannon/Simpson indices) in XJ samples. Significant compositional differences were observed between aerial (leaves) and underground (roots/pods) organs. Geographic location also markedly shaped endophytic communities, with stronger effects in roots and pods than in leaves—a pattern supported by PCoA combined with ANOSIM (R (roots) = 1, R (pods) = 0.874, R (leaves) = 0.336, respectively, p < 0.001). Saline–alkali adaptation led to a marked enrichment of Novosphingobium in roots and pods and of Halomonas in leaves compared to non-saline–alkali-grown peanuts. Furthermore, the endophytic communities within the same organ type varied significantly across the three saline–alkali sites. Redundancy analysis (RDA) identified the key environmental factors shaping bacterial community composition in the root samples from each location: available phosphorus (AP) and sulfate (SO₄²⁻) were the strongest predictors in XJ; available potassium (AK) and chloride (Cl⁻) in DY; and hydrolyzed nitrogen (HN), pH, soil organic matter (SOM), and bicarbonate (HCO₃⁻) in JL. These findings demonstrate that niches and geographical conditions determined the composition and relative abundance of endophytic bacteria in salt-tolerant peanuts, providing new insights into microbial ecological adaptation in saline–alkali ecosystems. Full article

Investigating the compositional characteristics of rhizosphere soil bacterial communities of Chinese seabuckthorn across different distribution areas and their relationship with habitat factors holds significant value for the development and utilization of characteristic medicinal plant resources in the Qinghai–Tibet Plateau. This study targeted rhizosphere soil from 12 distribution areas of Chinese seabuckthorn on the Qinghai–Tibet Plateau. By integrating measurements of soil and climatic parameters with high-throughput sequencing and redundancy analysis (RDA), the research systematically elucidated the characteristics of rhizosphere bacterial communities and their environmental driving mechanisms. The results revealed that rhizosphere bacterial communities of Chinese seabuckthorn across all 12 distribution areas were dominated by Proteobacteria, Acidobacteriota, and Actinobacteriota. Among them, the R2 habitat exhibited the highest amplicon sequence variant counts (3496), while R1, R8, R9, and R12 habitats showed significantly lower counts. α/β diversity analysis revealed that Shannon, Ace, and Chao1 indices in R2 and R7 habitats were significantly higher than those in R1 and R8. Regarding community aggregation patterns, soil bacterial communities in R1, R9, and R12 habitats exhibited the highest aggregation, while those in R2 and R11 habitats showed relatively lower aggregation. Functional prediction demonstrated that Metabolism dominated across all distribution areas (50.40–52.02%), with the R11 habitat exhibiting exceptionally high metabolic function abundance (>9300). Clustering analysis partitioned the 12 habitats into two distinct groups: one comprising R2, R6, R7, R9, R10, and R11, and the other containing the remaining six habitats. Redundancy analysis (RDA) further clarified that habitat factors, including altitude, soil water content (SWC), east longitude (EAST), and pH, were key drivers shaping bacterial community structure. This study underscores the pivotal regulatory role of environmental factors in shaping rhizosphere microbial diversity, community structure, and functional profiles of Chinese seabuckthorn, thereby providing a valuable scientific foundation for the sustainable development of characteristic medicinal plant resources on the Qinghai–Tibet Plateau. Full article

Integrated aquaculture, centered around polyculture involving multiple species, is a typical practice for the sustainable development of the aquaculture industry, capable of enhancing resource utilization efficiency, environmental stability, and overall productivity through establishing symbiotic interactions among species. This study employed multi-amplicon high-throughput sequencing to assess the ecological impacts of two polyculture methods involving river crabs on sediment bacteria, fungi, and protists. One method involved polyculturing river crabs with mandarin fish, silver carp, and the stone moroko (SPC), and the other involved polyculturing river crabs with only mandarin fish and silver carp (SMC). The results showed that, compared to the SMC group, the SPC group remarkably increased the Chao1 index of bacterial communities in pond sediment and decreased the Pielou_J index of protists. The relative abundances of all fungal phyla and most dominant bacterial and protistan phyla (top 10 in relative abundance) in the SPC group were considerably different from those in the SMC group. In the co-occurrence networks of bacterial, fungal, and protistan communities, the numbers of edges and nodes were higher in the SPC group than in the SMC group, and the habitat niche breadth of bacterial community was also notably increased in the SPC group. The levels of total carbon (TC), total nitrogen (TN), and phosphates within pond sediment in the SPC group were obviously lower than those in the SMC group, and were significantly correlated with the microbial communities, with TC being identified as the primary contributor driving changes in the microbial communities. All the findings collectively demonstrate that the polyculture of river crabs with mandarin fish, silver carp, and the stone moroko enhances the stability of bacterial, fungal, and protistan communities in sediment and enhances resource utilization efficiency in aquaculture, thereby preventing the environmental risks associated with excessive nutrient accumulation in sediment. Polyculture systems integrating river crabs with mandarin fish, silver carp, and the stone moroko represent a sustainable aquaculture model with significant ecological benefits. Full article

Activated sludge microorganisms in sewage treatment plants are crucial for controlling water pollution and protecting public health and the ecological environment. Activated sludge must have biodegradation, easy sedimentation, and separation functions. Filamentous bacteria play an essential role in floc formation and structure. However, low temperature, low load and low dissolved oxygen (DO) will destroy the balance between beneficial structural action and harmful overgrowth. In this study, the high-throughput sequencing (HTS) dataset of 16s rRNA gene sequence V3–V4 amplicons from 30 activated sludge samples from the Chuanhu Sewage Treatment Plant in Changchun was analyzed to investigate the abundance distribution of filamentous bacteria and further determine the main operating parameters and environmental factors. The experimental results showed that the filamentous bacterial community accounted for a large part of the entire microbial community, with the total filamentous bacterial percentage in each sample ranging from 7.32% to 56.81%, with large fluctuations in abundance and consistent with the SVI value. Although most of them were in flocs, they occasionally caused sedimentation problems when the water temperature was low. With 14 species of filamentous bacteria detected, the population structure of filamentous bacteria in the thermophilic, variable-temperature and low-temperature periods was universal and specific. The groups with a detection frequency of 100%, high abundance, and significant fluctuations in distribution were Microthrix parvicella and Nostocoida limicola I. The Pearson correlation analysis showed that the total abundance of filamentous bacteria and the fluctuation distribution of dominant filamentous bacteria abundance were significantly correlated with water temperature, sludge load, sludge age, and mixed liquid suspended solids (MLSS). Full article

Increasing demand for high-quality food is driving the development of biologized farming methods, which involve the use of microorganisms, including endophytes, to stimulate plant growth. However, research on the composition of endosphere microbiomes is limited. The study presents an analysis of the bacterial endophytic microbiome in lettuce seeds (Lactuca sativa L., cv. Ozornik) using high-throughput sequencing of 16S rRNA amplicons. It evaluates the taxonomic composition and putative functional properties of seed endophytic bacteria. The microbial community exhibited low diversity (Shannon index ranged from 1.1 to 1.84, Simpson index from 0.57 to 0.83). The bacterial endophytic community of lettuce seeds was dominated by Pseudomonadota (83%), Actinomycetota (14%), and Bacillota (3%). The genera identified within the microbiome included Pantoea (32%), Rhodococcus (13%), Candidatus Profftella (13%), Janthinobacterium (7%), Pseudomonas (9%), Enterococcus (3%), and Alcaligenes (2%), which exhibit a broad spectrum of beneficial properties: plant growth promotion (PGPB), suppression of phytopathogens, enhanced stress tolerance, participation in contaminant biodegradation, and heavy metal detoxification. The structure and functional potential of the microbiome vary between samples, potentially due to differences in source material and cultivation conditions. The obtained results expand our understanding of the composition and functions of endophytic bacteria in lettuce seeds, which is important for the development of novel biocontrol agents for plants consumed by humans in an unprocessed form. Full article

Rhizosphere microbiota are key drivers of plant nutrition, immunity, and stress tolerance. Abies ziyuanensis L. K. Fu & S. L. Mo (Pinaceae) is an endangered conifer endemic to China, and its persistence may depend on its interactions with its belowground microbiome. However, how soil-borne bacterial functional groups respond to, and potentially support, A. ziyuanensis remains unclear. Based on amplicon high-throughput sequencing data of the 16S rRNA gene and soil physicochemical properties, the bacterial community structure in the rhizosphere soil of A. ziyuanensis in Yinzhu Laoshan National Nature Reserve in Guangxi Zhuang Autonomous Region, China, was analyzed, and the potential ecological functions and phenotypic characteristics of the bacterial community were predicted to determine the functional taxa characteristics (nitrogen cycle, phototrophy, and chemoheterotrophy) and dominant soil environmental factors. Proteobacteria, Acidobacteria, Actinobacteria, Planctomycetes, Verrucomicrobia, and Chloroflexi were the dominant bacterial taxa in the A. ziyuanensis rhizosphere soil, and all bacteria were significantly positively correlated with soil NO₃⁻-N (R = 0.47, p = 0.0079). Based on FAPROTAX, the A. ziyuanensis rhizosphere soil bacterial community had chemoheterotrophic-related functions, which were more prevalent than nitrogen cycle- and phototrophic-related functions, and the relative abundance of bacteria with nitrogen cycle-related functions was higher than that of those with phototrophic functions. The nitrogen nutrient- and phototrophic-related functional taxa in the rhizosphere soil bacterial community had significant correlations with soil physicochemical properties, whereas the chemoheterotrophic-related functional taxa did not show a significant correlation. Based on BugBase phenotype prediction, Acidobacteria, Proteobacteria, and Chloroflexi made the greatest contribution to the phenotype, with pathogenic and stress tolerance being the most important phenotypes. The pathogenic and stress-tolerant bacteria all belonged to Proteobacteria. The rhizosphere bacteria exhibited rich diversity and dominated several biogeochemical cycling processes. This study identifies beneficial rhizosphere bacteria of A. ziyuanensis, providing a theoretical basis for conserving soil bacterial diversity and guiding the targeted recruitment of functional bacteria by the endangered plant. Full article

Antheraea pernyi (Lepidoptera: Saturniidae) is an economically important silk-producing insect, whose gut microbiota play a crucial role in growth, development, and nutrient metabolism. This study focused on the entire larval developmental stages of A. pernyi. Using the Illumina MiSeq high-throughput sequencing platform, we performed 16S rRNA gene amplicon sequencing on the gut microbiota of laboratory-reared A. pernyi larvae, analyzing in detail the composition and diversity characteristics of the gut microbial communities across all five instars (1st to 5th instar). Additionally, functional predictions were conducted to explore the potential roles of these microbiota during larvae development. The study revealed that the core gut microbiota of A. pernyi larvae primarily consisted of Actinomycetota (39.78%), Cyanobacteriota (32.46%), Bacillota (18.08%), and Pseudomonadota (9.02%). Among these, Actinomycetota dominated in the 1st to 4th-instar larvae, while Cyanobacteriota became the predominant phylum in the 5th instar. Linear discriminant analysis effect size identified statistically significant biomarkers across different instar larvae of A. pernyi. Alpha diversity analysis showed that gut microbiota diversity initially increased and then decreased with larval development, peaking in the 3rd instar, and reaching its lowest level in the 5th instar. Principal coordinate analysis (PCoA) of beta diversity indicated that the gut microbiota structures of the 1st to 4th instars were similar but significantly differed from that of the 5th instar. Functional prediction analysis based on the KEGG database revealed that Carbohydrate metabolism and Amino acid metabolism-related genes were significantly lower in the 5th instar compared to other instars, while Energy metabolism and Cofactor and vitamin metabolism-related genes were significantly higher. This study offers valuable insights for the development of gut microbial resources in Lepidoptera insects. Full article

Fermentation is a crucial stage in the production of washed mild coffees, as it enables the generation of compounds that influence overall quality. The conditions to optimize this process are still unknown. This study evaluated the effects of fermenting coffee fruits and depulped coffee under two conditions: an open tank (semi-anaerobic-SA) and a closed tank (self-induced anaerobic fermentation, SIAF) over 192 h. Samples were taken every 24 h using a sacrificial bioreactor. A randomized complete block design with a factorial arrangement (2 × 2 + 1), plus a standard control, was employed, incorporating two factors: coffee type and fermentation condition. High-throughput sequencing of 16S and ITS amplicons identified an average of 260 ± 71 and 101 ± 24 OTUs, respectively. Weisella was the dominant lactic acid bacteria, followed by Leuconostoc and Lactiplantibacillus. Acetic acid bacteria, mainly Acetobacter, were more abundant under semi-anaerobic conditions. The yeast genera most affected by the fermentation condition were Pichia, Issatchenkia, and Wickerhamomyces. Repeated measures analysis revealed significant differences in pH, glucose consumption, lactic acid production, dry matter content, embryo viability, and the percentage of healthy beans. Principal component analysis was used to develop an index that integrates physical, physiological, and sensory quality variables, thereby clarifying the impact of each treatment. Samples from shorter fermentation times and SIAF conditions scored closest to 1.0, reflecting the most favorable outcomes. Otherwise, samples from longer fermentation times in both depulped and coffee fruits scored 0.497 and 0.369, respectively, on the SA condition. These findings support technically and economically beneficial fermentation strategies. Full article

Apricots are affected by many abiotic and biotic factors that could negatively impact their vitality and yield, leading to branch and tree dieback. Knowledge of the microbiome composition is key to choosing the optimal measurement strategy. The effect of the two different growing systems, i.e., organic (ORG) and integrated pest management (IPM), on the apricot fungal microbiome was studied. The inner bark was used to isolate DNA, and the present fungi were analyzed using a metagenomics high-throughput sequencing (HTS) profiling approach of the data obtained based on the Illumina sequencing of the ITS1-ITS2 amplicons of the 18S rRNA gene. Of the 20 analyzed samples, Ascomycota was the dominant phylum, and Dothiomycetes was the most abundant. Basidiomycota was the less frequent, with Tremellomycetes being the predominant within this phylum. PCA analysis showed the complete separation of the samples obtained from the orchards grown under the ORG and IPM systems. Cladosporia, Alternaria, Aureobasidium, and Visniacozyma were detected in all samples, but they dominated the IPM samples. Filobasiadiales were recognized as an indicator species for ORG management, while Caliciales, Lecanorales, Lichinales, Mycosphaerellales, Myriangiales, Phacidiales, Teloschistales, and Thelebolales were identified as indicator species for IPM management. Based on the order and genus levels, a significantly higher fungal microbiome richness was detected in the ORG samples. This could be connected to the environmentally beneficial growing system applied in the orchard, but it is impossible to assess the risk of trunk disease development or premature apricot tree decline. Full article

Hippophae rhamnoides subsp. sinensis Rousi has high ecological and medicinal value, and it is an important plant resource unique to the arid regions of Northwest China. Exploring the influence of climate characteristics and soil factors on the composition, diversity, and function of the rhizosphere bacterial community of Chinese seabuckthorn is of great value for developing and popularizing characteristic plant resources in the arid regions of Northwest China. In this study, the rhizosphere soil of 13 Chinese seabuckthorn distribution areas in the northwest of China was taken as the research object, the bacterial community map was constructed based on 16S rRNA gene high-throughput sequencing technology, and the species abundance composition, structural diversity, molecular co-occurrence network, and phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt), as well as the function of rhizosphere soil bacterial community, were systematically studied. Combined with Mantel test and redundancy analysis (RDA), the key habitat factors driving the rhizosphere soil bacterial community structure of Chinese seabuckthorn were explored. The results showed that: (1) The number of amplicon sequence variants (ASVs) in rhizosphere soil bacterial community of Chinese seabuckthorn was the highest in S2(3072) and the S12(3637), and the lowest in the S11(1358) and S13(1996). The rhizosphere soil bacterial community was primarily composed of Proteobacteria, Actinobacteriota, and Acidobacteriota. Except for the S6 and S11 habitats, the dominant bacterial genera were mainly Achromobacter, Acidobacter (RB41), and Sphingomonas. (2) The α and β diversity of rhizosphere soil bacterial communities of Chinese seabuckthorn across 13 distribution areas were significantly different. The number of operational taxonomic units (OTUs), Ace index, and Chao 1 index of soil bacterial community in the S12 distribution area are the highest, and they are the lowest in S11 distribution area, with significant differences. The aggregation of soil bacterial communities in the S5 and S10 distribution areas is the highest, while it is the lowest in the S6 and S11 distribution areas. (3) PICRUSt function classification of soil bacteria showed that Metabolism and Genetic Information Processing functions were the strongest across all distribution areas, with S10 exhibiting higher functional capacity than other areas and S11 showing the weakest. (4) Cluster analysis revealed that soil bacteria across the 13 distribution areas were clustered into two groups, with S10 and S12 distribution areas as one group (Group 1) and the remaining 11 distribution areas as another group (Group 2). (5) Redundancy analysis revealed that pH was the key soil environmental factor driving the rhizosphere soil bacterial community α-diversity of Chinese seabuckthorn, followed by altitude (ALT) and soil water content (SWC). In summary, Chinese seabuckthorn prefers neutral to alkaline soils, and environmental factors play an important role in driving bacterial diversity, community structure, functional profiles, and co-occurrence networks in rhizosphere soil of Chinese seabuckthorn. Full article

In response to the challenges posed by soil degradation in the arid regions of Xinjiang, China, green and organic management practices have emerged as effective alternatives to conventional agricultural management methods, helping to mitigate soil degradation by promoting natural soil recovery and ecological balance. However, most of the existing studies focus on a single management practice or indicator and lack a systematic assessment of the effects of integrated orchard management in arid zones. This study aims to investigate how different agricultural management practices influence soil physicochemical properties and inter-root microbial communities in apple orchards in Xinjiang and to identify the main physicochemical factors affecting the composition of inter-root microbial communities. Inter-root soil samples were collected from apple orchards under green management (GM), organic management (OM), and conventional management (CM) in major apple-producing regions of Xinjiang. Microbial diversity and community composition of the samples were analyzed using high-throughput amplicon sequencing. The results revealed significant differences (p < 0.05) in soil physicochemical properties across different management practices. Specifically, GM significantly reduced soil pH and C:N compared with OM. Both OM and GM significantly decreased soil available nutrient content compared with CM. Moreover, GM and OM significantly increased bacterial diversity and changed the community composition of bacteria and fungi. Proteobacteria and Ascomycota were identified as the dominant bacteria and fungi, respectively, in all management practices. Linear discriminant analysis (LEfSe) showed that biomarkers were more abundant under OM, suggesting that OM may contribute to ecological functions through specific microbial taxa. Co-occurrence network analysis (building a network of microbial interactions) demonstrated that the topologies of bacteria and fungi varied across different management practices and that OM increased the complexity of microbial co-occurrence networks. Mantel test analysis (analyzing soil factors and microbial community correlations) showed that C:N and available potassium (AK) were significantly and positively correlated with the community composition of bacteria and fungi, and that C:N, soil organic carbon (SOC), and alkaline hydrolyzable nitrogen (AN) were significantly and positively correlated with the diversity of fungi. Redundancy analysis (RDA) further indicated that SOC, C:N, and AK were the primary soil physicochemical factors influencing the composition of microbial communities. This study provides theoretical guidance for the sustainable management of orchards in arid zones. 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