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Identification of bioactives from the red seaweed Asparagopsis taxiformis that promote antimethanogenic activity in vitro

Abstract

Asparagopsis taxiformis has potent antimethanogenic activity as a feed supplement at 2 % of organic matter in in vitro bioassays. This study identified the main bioactive natural products and their effects on fermentation using rumen fluid from Bos indicus steers. Polar through to non-polar extracts (water, methanol, dichloromethane and hexane) were tested. The dichloromethane extract was most active, reducing methane production by 79 %. Bromoform was the most abundant natural product in the biomass of Asparagopsis (1723 μg g−1 dry weight [DW] biomass), followed by dibromochloromethane (15.8 μg g−1 DW), bromochloroacetic acid (9.8 μg g−1 DW) and dibromoacetic acid (0.9 μg g−1 DW). Bromoform and dibromochloromethane had the highest activity with concentrations ≥1 μM inhibiting methane production. However, only bromoform was present in sufficient quantities in the biomass at 2 % organic matter to elicit this effect. Importantly, the degradability of organic matter and volatile fatty acids were not affected at effective concentrations.

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Acknowledgments

This research is supported by funding from the Australian Government Department of Agriculture, the Australian Government through the Australian Renewable Energy Agency, and the Advanced Manufacturing Cooperative Research Centre (AMCRC), funded through the Australian Government’s Cooperative Research Centre Scheme. We thank Dr. Mike Devery for kindly supplying the bromochloromethane used in this study. We also thank Pedro Henrique de Paula Silva for reviewing the manuscript.

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Correspondence to Lorenna Machado.

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Rumen fluid was collected from four rumen-fistulated Brahman (B. indicus) steers, which were maintained according to guidelines approved by CSIRO Animal Ethics Committee (A5/2011) and in accordance with the Australian Code of Practice for the Care and Use of Animals for Scientific Purposes.

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Figure S1
figure5

Effects of Asparagopsis biomass and extracts (doses equivalent to Asparagopsis 2 % OM) on production of (A) total gas (TGP), and (B) CH4 over a 24 h fermentation period. Error bars represent ± SE (n = 3). DCM = dichloromethane. (GIF 73 kb)

High Resolution image (TIF 284 kb)

Table S1

Results of full factorial permutational analyses of variance (PERMANOVAs) testing the effects of the fixed factors pure compound, and assay-concentration on total gas production (TGP), methane (CH4), and hydrogen (H2). Analyses were conducted in Primer v6 (Primer-E Ltd, UK) using Bray-Curtis dissimilarities on fourth root transformed data and 9999 unrestricted permutations of raw data. Pseudo F (F) and P values are presented (n = 3). (DOCX 14 kb)

Table S2

ANOVA results of the natural product yields of each solvent extract of Asparagopsis biomass [μg g-1 dry weight (DW)], (Experiment 2). (DOCX 14 kb)

Table S3

Results of full factorial permutational analyses of variance (PERMANOVAs) testing the effects of the fixed factors pure compound, and assay-concentration on total gas production (TGP), methane (CH4), and hydrogen (H2). Analyses were conducted in Primer v6 (Primer-E Ltd, UK) using Bray-Curtis dissimilarities on fourth root transformed data and 9999 unrestricted permutations of raw data. Pseudo F (F) and P values are presented (n = 3). (DOCX 14 kb)

Table S4

Results of full factorial permutational analyses of variance (PERMANOVAs) testing the effects of the fixed factors pure compound and assay-concentration on total gas production (TGP), methane (CH4), and hydrogen (H2), (Experiment 3). Analyses were conducted in Primer v6 (Primer-E Ltd, UK) using Bray-Curtis dissimilarities on fourth root transformed data and 9999 unrestricted permutations of raw data (n = 4). (DOCX 18 kb)

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Machado, L., Magnusson, M., Paul, N.A. et al. Identification of bioactives from the red seaweed Asparagopsis taxiformis that promote antimethanogenic activity in vitro. J Appl Phycol 28, 3117–3126 (2016). https://doi.org/10.1007/s10811-016-0830-7

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Keywords

  • Red macroalgae
  • Rhodophyta
  • Seaweed
  • Natural products
  • Methane inhibitors
  • Bromoform