Cited by 147Ruminant Nutrition and Digestive PhysiologyAnaerobic Digestion and Biogas ProductionGut microbiota and health

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Study of methanogen communities associated with different rumen protozoal populations

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Belanche, Alejandro, de la Fuente, Gabriel, Newbold, Charles J.·2014·FEMS Microbiology Ecology

DOI 10.1111/1574-6941.12423PMC PMC4286163

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Abstract

Protozoa-associated methanogens (PAM) are considered one of the most active communities in the rumen methanogenesis. This experiment investigated whether methanogens are sequestrated within rumen protozoa, and structural differences between rumen free-living methanogens and PAM. Rumen protozoa were harvested from totally faunated sheep, and six protozoal fractions (plus free-living microorganisms) were generated by sequential filtration. Holotrich-monofaunated sheep were also used to investigate the holotrich-associated methanogens. Protozoal size determined the number of PAM as big protozoa had 1.7-3.3 times more methanogen DNA than smaller protozoa, but also more endosymbiotic bacteria (2.2- to 3.5-fold times). Thus, similar abundance of methanogens with respect to total bacteria were observed across all protozoal fractions and free-living microorganisms, suggesting that methanogens are not accumulated within rumen protozoa in a greater proportion to that observed in the rumen as a whole. All rumen methanogen communities had similar diversity (22.2 ± 3.4 TRFs). Free-living methanogens composed a conserved community (67% similarity within treatment) in the rumen with similar diversity but different structures than PAM (P < 0.05). On the contrary, PAM constituted a more variable community (48% similarity), which differed between holotrich and total protozoa (P < 0.001). Thus, PAM constitutes a community, which requires further investigation as part of methane mitigation strategies.

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Figure 1 Diagram depicting sheep inoculation and rumen sampling.CC BY-NC · PubMed Central

Figure 2 Fluorescence microscopy images of the different protozoal fractions using propidium iodide dye and rhodamine filters. Protozoa fractions were obtained from holotrich-monofaunated (H60 and H20) and totally faunated sheep (F80, F60, F45, F35, F20, F5 and F < 5) using different nylon meshes (80, 60, 45, 35, 20 and 5 μm pore size).CC BY-NC · PubMed Central

Figure 3 (a) Ratio bacteria/protozoa, (b) methanogens/protozoa and (c) methanogens/bacteria in different rumen protozoal fractions obtained from holotrich-monofaunated (H60 and H20) and totally faunated sheep (F80, F60, F45, F35, F20, F5 and F < 5) using different nylon meshes (80, 60, 45, 35, 20 and 5 μm pore size). Data were log10-transformed to attain normality. Bars with different letters (a, b, c, d, e) of the same colour differ (P < 0.05).CC BY-NC · PubMed Central

Figure 4 (a) PCoA illustrating the differences in the endosymbiotic methanogens associated with different rumen protozoal fractions obtained from holotrich-monofaunated (H) and totally faunated sheep (F). Big circles indicate the 90% confidential interval. (b) Dendrogram depicting the effect of total protozoa fractionation on their endosymbiotic methanogen populations. Protozoa fractions (F80, F60, F45, F35, F20, F5 and F < 5) were generated by a sequential filtration of rumen fluids from different sheep (A, B, C and D) through nylon meshes with a pore size of 80, 60, 45, 35, 20 and 5 μm, respectively.CC BY-NC · PubMed Central

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Authors: Belanche, A., de la Fuente, G., & Newbold, C. J.
Affiliations: Aberystwyth University (GB)
Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, Aberystwyth, UK
Corresponding author: Newbold, Charles J.
ORCID: Belanche, Alejandro
Newbold, Charles J.
Venue: FEMS Microbiology Ecology
Publisher: Oxford University Press (OUP)
Published: 2014-12-01
Volume: 90
Issue: 3
Pages: 663-677
ISSN: 0168-6496 (ISSN-L)
1574-6941
DOI: 10.1111/1574-6941.12423
PMC: PMC4286163
OpenAlex: W2018339579
Cited by: 147
References: 60
Citations by year: 2026: 6 · 2025: 16 · 2024: 20 · 2023: 11 · 2022: 11 · 2021: 19 · 2020: 12 · 2019: 16 · 2018: 10 · 2017: 7 · 2016: 10 · 2015: 8 · 2014: 1
FWCI: 17.27
Type: journal article
Language: en
Open access: bronze
doi.org
Locations: FEMS Microbiology Ecology (publishedVersion)
Europe PMC (PubMed Central) (publishedVersion, cc-by-nc)
PubMed Central (publishedVersion, cc-by-nc)
PubMed (publishedVersion)
RECERCAT (Consorci de Serveis Universitaris de Catalunya) (publishedVersion, cc-by-nc)
Repositori ObertUDL (University of Lleida) (publishedVersion, other-oa)
LA Referencia (Red Federada de Repositorios Institucionales de Publicaciones Científicas) (submittedVersion, cc-by-nc)
Funders: European Commission (FP7, REDNEX project)
Topics: Life Sciences › Agricultural and Biological Sciences › Animal Science and Zoology › Ruminant Nutrition and Digestive Physiology
Physical Sciences › Energy › Renewable Energy, Sustainability and the Environment › Anaerobic Digestion and Biogas Production
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gut microbiota and health
Keywords: rumen methanogenesis, rumen protozoa, endosymbiotic bacteria, methane mitigation, sheep
MeSH terms: Animals, Bacteria, Methane, Polymorphism, Restriction Fragment Length, Rumen, Symbiosis, Trichostomatida, Euryarchaeota, Sheep, Domestic

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Study of methanogen communities associated with different rumen protozoal populations

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