Ruminant livestock are important sources of human food and global greenhouse gas emissions. Feed degradation and methane formation by ruminants rely on metabolic interactions between rumen microbes and affect ruminant productivity. Rumen and camelid foregut microbial community composition was determined in 742 samples from 32 animal species and 35 countries, to estimate if this was influenced by diet, host species, or geography. Similar bacteria and archaea dominated in nearly all samples, while protozoal communities were more variable. The dominant bacteria are poorly characterised, but the methanogenic archaea are better known and highly conserved across the world. This universality and limited diversity could make it possible to mitigate methane emissions by developing strategies that target the few dominant methanogens. Differences in microbial community compositions were predominantly attributable to diet, with the host being less influential. There were few strong co-occurrence patterns between microbes, suggesting that major metabolic interactions are non-selective rather than specific.

The rumen is a complex ecosystem composed of anaerobic bacteria, protozoa, fungi, methanogenic archaea and phages. These microbes interact closely to breakdown plant material that cannot be digested by humans, whilst providing metabolic energy to the host and, in the case of archaea, producing methane. Consequently, ruminants produce meat and milk, which are rich in high-quality protein, vitamins and minerals, and therefore contribute to food security. As the world population is predicted to reach approximately 9.7 billion by 2050, an increase in ruminant production to satisfy global protein demand is necessary, despite limited land availability, and whilst ensuring environmental impact is minimized. Although challenging, these goals can be met, but depend on our understanding of the rumen microbiome. Attempts to manipulate the rumen microbiome to benefit global agricultural challenges have been ongoing for decades with limited success, mostly due to the lack of a detailed understanding of this microbiome and our limited ability to culture most of these microbes outside the rumen. The potential to manipulate the rumen microbiome and meet global livestock challenges through animal breeding and introduction of dietary interventions during early life have recently emerged as promising new technologies. Our inability to phenotype ruminants in a high-throughput manner has also hampered progress, although the recent increase in “omic” data may allow further development of mathematical models and rumen microbial gene biomarkers as proxies. Advances in computational tools, high-throughput sequencing technologies and cultivation-independent “omics” approaches continue to revolutionize our understanding of the rumen microbiome. This will ultimately provide the knowledge framework needed to solve current and future ruminant livestock challenges.

Significance and Impact of the Study: We investigated, over a 24-h period in sacco, whether attachment of rumen microbiota to perennial ryegrass (PRG) showed successional changes in diversity. Knowledge of the bacterial species that attach to PRG over time may aid our understanding of the temporal function of the attached microbiota and ultimately permit the development of novel strategies for improving animal production to meet the future demands for meat and milk. AbstractThis study investigated successional colonization of perennial ryegrass (PRG) by the rumen microbiota. PRG grown for 6 weeks in a greenhouse was incubated in sacco in the rumens of three Holstein 9 Freisian cows over a period of 24 h. PRG incubated within the rumen was subsequently harvested at various time intervals postincubation to assess colonization over time. DGGE-based dendograms revealed the presence of distinct primary (0-2 h) and secondary (4 h onwards) attached bacterial communities. Moving window analysis, band number and Shannon-Wiener diversity indices suggest that after 2 h a proportion of primary colonizing bacteria detach, to be replaced with a population of secondary colonizing bacteria between 2 and 4 h after entry of PRG into the rumen. Sequencing and classification of bands lost and gained between 2 and 4 h showed that the genus Prevotella spp. was potentially more prevalent following 4 h of incubation, and Prevotella spp. 16S rDNA-based QPCR supported this finding somewhat, as 2-to 4-h Prevotella QPCR data were greater but not significantly so. Low-temperature scanning electron microscopy showed that attached bacteria were predominantly enveloped in extracellular polymeric substances. In conclusion, colonization of fresh PRG is biphasic with primary colonization completed within 2 h and secondary colonization commencing after 4 h of attachment in this study.

Understanding the relationship between ingested plant material and the attached microbiome is essential for developing methodologies to improve ruminant nutrient use efficiency. We have previously shown that perennial ryegrass (PRG) rumen bacterial colonization events follow a primary (up to 4 h) and secondary (after 4 h) pattern based on the differences in diversity of the attached bacteria. In this study, we investigated temporal niche specialization of primary and secondary populations of attached rumen microbiota using metagenomic shotgun sequencing as well as monitoring changes in the plant chemistry using mid-infrared spectroscopy (FT-IR). Metagenomic Rapid Annotation using Subsystem Technology (MG-RAST) taxonomical analysis of shotgun metagenomic sequences showed that the genera Butyrivibrio, Clostridium, Eubacterium, Prevotella, and Selenomonas dominated the attached microbiome irrespective of time. MG-RAST also showed that Acidaminococcus, Bacillus, Butyrivibrio, and Prevotella rDNA increased in read abundance during secondary colonization, whilst Blautia decreased in read abundance. MG-RAST Clusters of Orthologous Groups (COG) functional analysis also showed that the primary function of the attached microbiome was categorized broadly within “metabolism;” predominantly amino acid, carbohydrate, and lipid metabolism and transport. Most sequence read abundances (51.6, 43.8, and 50.0% of COG families pertaining to amino acid, carbohydrate and lipid metabolism, respectively) within these categories were higher in abundance during secondary colonization. Kyoto encyclopedia of genes and genomes (KEGG) pathways analysis confirmed that the PRG-attached microbiota present at 1 and 4 h of rumen incubation possess a similar functional capacity, with only a few pathways being uniquely found in only one incubation time point only. FT-IR data for the plant residues also showed that the main changes in plant chemistry between primary and secondary colonization was due to increased carbohydrate, amino acid, and lipid metabolism. This study confirmed primary and secondary colonization events and supported the hypothesis that functional changes occurred as a consequence of taxonomical changes. Sequences within the carbohydrate metabolism COG families contained only 3.2% of cellulose activities, on average across both incubation times (1 and 4 h), suggesting that degradation of the plant cell walls may be a key rate-limiting factor in ensuring the bioavailability of intra-plant nutrients in a timely manner to the microbes and ultimately the animal. This suggests that a future focus for improving ruminant nutrient use efficiency should be altering the recalcitrant plant cell wall components and/or improving the cellulolytic capacity of the rumen microbiota.

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Huws, S. A., Mayorga, O. L., Theodorou, M. K., Kim, J. S., Cookson, A., Newbold, J., Kingston-Smith, A. H. (2014). Differential Colonization of Plant Parts by the Rumen Microbiota is likely to be due to Different Forage Chemistries. Journal of Microbial and Biochemical Technology, 6 (2), 80-86In this experiment we investigated the hypothesis that heterogeneity of plant structures presents disparity in niches available for colonisation by the rumen microbiota resulting in differential colonisation. Fresh perennial ryegrass (PRG) stem and leaves were incubated in the presence of rumen bacteria under rumen-like conditions with incubations harvested at many time intervals up to 24 h. In vitro dry matter degradability (IVDMD) of stem material was lower than that of leaves at all harvesting times. Denaturing Gradient Gel Electrophoresis (DGGE) derived dendrograms, canonical analysis of principal coordinates (CAP) and PERMANOVA demonstrated that diversity of bacteria attached to PRG stem and leaf material was different at all harvesting times although QPCR data showed similar quantities of bacterial 16S rDNA on stem and leaf material at all harvesting times. Conversely, bacterial diversity on abaxial and adaxial leaf surfaces was similar, but 16S rDNA quantity differed with more 16S rDNA on the adaxial surface at all harvesting times. Image analysis of low temperature scanning electron microscopy (LTSEM) pictures confirmed that biofilm coverage on the adaxial surface was greater than the abaxial surface. We demonstrate that differing plant parts can affect attached bacterial diversity and/or 16S rDNA quantity present. This result is consistent with concepts of niche specialisation by the rumen microbiota. This observation is particularly relevant to understanding rumen plant-microbe interactions which is necessary for development of novel strategies for improving ruminant nutrient use efficiency.publishersversionPeer reviewe

Ruminant livestock are important sources of human food and global greenhouse gas emissions. Feed degradation and methane formation by ruminants rely on metabolic interactions between rumen microbes and affect ruminant productivity. Rumen and camelid foregut microbial community composition was determined in 742 samples from 32 animal species and 35 countries, to estimate if this was influenced by diet, host species, or geography. Similar bacteria and archaea dominated in nearly all samples, while protozoal communities were more variable. The dominant bacteria are poorly characterised, but the methanogenic archaea are better known and highly conserved across the world. This universality and limited diversity could make it possible to mitigate methane emissions by developing strategies that target the few dominant methanogens. Differences in microbial community compositions were predominantly attributable to diet, with the host being less influential. There were few strong co-occurrence patterns between microbes, suggesting that major metabolic interactions are non-selective rather than specific.

Rumen fermentation and methanogenesis are vital metabolic processes in cattle and are carried out by microbial populations that are affected by dietary factors such as secondary metabolites, nutritional composition and degradability. The aim of this study was to monitor populations of total bacteria, total methanogens and Butirivibrio fibrisolvens in the rumen of Lucerne heifers fed on diets typical of intensive silvopastoral systems (ISS) or of a traditional (control) system. Rumen contents (RC) were collected orally from eight heifers consuming 100% Cynodon plectostachyus (control) and 76% C. plectostachyus + 24% Leucaena leucocephala (ISS) following a crossover design and DNA was extracted and quantified by quantitative PCR. Populations [Log10 (ng g-1 RC)] were 5.6 and 5.8 for total bacteria (p = 0.5343), 3.6 and 3.5 for B. fibrisolvens (p = 0.4742) and 5.0 and 5.3 for total methanogens (p = 0.2661) respectively in control and ISS diets. However, when measured in a separate parallel study, enteric methane emissions (g kg-1 of fermented dry matter) were significantly reduced with the inclusion of L. leucocephala. This fact indicated the importance of investigating the structure, function and interactions of populations beyond quantitative analysis to determine how diet affects rumen microbial populations and their function.