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In Vitro Response of Rumen Microbiota to the Antimethanogenic Red Macroalga Asparagopsis taxiformis

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Abstract

The red macroalga Asparagopsis taxiformis has been shown to significantly decrease methane production by rumen microbial communities. This has been attributed to the bioaccumulation of halogenated methane analogues produced as algal secondary metabolites. The objective of this study was to evaluate the impact of A. taxiformis supplementation on the relative abundance of methanogens and microbial community structure during in vitro batch fermentation. Addition of A. taxiformis (2% organic matter) or the halogenated methane analogue bromoform (5 μM) reduced methane production by over 99% compared to a basal substrate-only control. Quantitative PCR confirmed that the decrease in methane production was correlated with a decrease in the relative abundance of methanogens. High-throughput 16S ribosomal RNA gene amplicon sequencing showed that both treatments reduced the abundance of the three main orders of methanogens present in ruminants (Methanobacteriales, Methanomassiliicoccales and Methanomicrobiales). Shifts in bacterial community structure due to the addition of A. taxiformis and 5 μM bromoform were similar and concomitant with increases in hydrogen concentration in the headspace of the fermenters. With high potency and broad-spectrum activity against rumen methanogens, A. taxiformis represents a promising natural strategy for reducing enteric methane emissions from ruminant livestock.

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Acknowledgements

We thank Dr. Mike Devery for kindly supplying the bromochloromethane used in this study and Dr. Matthew Vucko for assistance with the experiments. We also thank Jeffrey Palpratt from the College of Public Health, Medical & Veterinary Sciences for handling and maintenance of the donor steers, and Dr. Shane Askew from the Advanced Analytical Centre, JCU, for analytical advice.

Funding

This research was supported by the Australian Government Department of Agriculture and Meat & Livestock Australia, funded as part of the National Livestock Methane Program. It was also supported by MBD Energy and the Australian Government through the Advanced Manufacturing Cooperative Research Centre, funded through the Australian Government’s Cooperative Research Centre Scheme.

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Correspondence to Lorenna Machado.

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Experimental protocols were approved by the CSIRO Animal Ethics Committee (A5/2011) under the Australian Code of Practice for Care and Use of Animals for Scientific Purposes

Electronic supplementary material

Table S1

Mean gas production parameters after 72 h incubation (n = 3) (DOCX 13 kb)

Table S2

OTU table with FASTA sequences and taxonomic lineages for experimental and technical replicates. Samples are labelled as treatment_time.Nx, where N is 1, 2 or 3 to denote the experimental replicate; and x is a or b to denote the technical replicate. OTU labelling corresponds to sequences deposited in NCBI Genbank under accession numbers KT168398 – KT174433. (XLSX 1135 kb)

Fig. S1

Principal co-ordinate analysis plot showing relationships between microbial communities under different experimental conditions, based on unweighted UniFrac metric of β diversity. BF1: 1 μM bromoform; BF5: 5 μM bromoform Asp: 2% A. taxiformis; BCM: 5 μM bromochloromethane. (GIF 85 kb)

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Machado, L., Tomkins, N., Magnusson, M. et al. In Vitro Response of Rumen Microbiota to the Antimethanogenic Red Macroalga Asparagopsis taxiformis . Microb Ecol 75, 811–818 (2018). https://doi.org/10.1007/s00248-017-1086-8

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