Journal Articles
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Item Isolation and characterization of Methanosphaera sp. ISO3-F5, a member of a novel and widespread species of rumen methanogens growing with methanol plus hydrogen(The Microbes, 2024-12-03) Jeyanathan J; Palevich N; Reilly K; Palevich FP; Maclean PH; Li D; Altermann E; Kim CC; van Scheepstal IM; Hoskin SO; Kelly WJ; Leahy SC; Attwood GT; Ronimus RS; Henderson G; Janssen PHRumen methanogens predominantly fall into two physiological groups: hydrogenotrophs which use hydrogen (H2) to reduce carbon dioxide (CO2) to methane (CH4), and methylotrophs which use H2 to reduce methanol and methylamines as substrates for methanogenesis. We used a dilution to extinction approach to isolate two hydrogenotrophic Methanocatella spp. and four cultures of methylotrophic methanogens from sheep rumen contents. Three of the methylotrophs were stable mixed cultures containing methanogens belonging to different lineages of the order Methanomassiliicoccales and one was a pure Methanosphaera culture. Methanosphaera sp. ISO3-F5 has a comparatively large genome (2.68 Mb) comprised of two replicons, a chromosome and a megaplasmid. The genome has an average G + C content of 30.5 % and encodes 2360 putative protein-coding genes. Cells of ISO3-F5 have a spherical shape, 0.6–1.2 µm in diameter, usually occurring in pairs or loose clumps, and have no flagellum. Cells stain Gram positive, have a single thick cell wall and divide by the formation of a cross wall. The optimum temperature for growth was 39°C to 42°C and the optimum pH was 6.7–6.8. Acetate was required for growth, but CH4 was not produced from acetate, formate, ethanol, methylamine, or isopropanol with or without H2/CO2. Volatile fatty acids and rumen fluid were also found to enhance the growth of ISO3-F5, while coenzyme M did not. ISO3-F5 produced CH4 from methanol in the presence of H2 and the genes encoding the necessary methanogenesis pathway have been identified. Based on morphological, physiological, and genomic characteristics, ISO3-F5 is a new species of the genus Methanosphaera. Our study shows that simple isolation methods allowed us to culture diverse and significant members of the rumen methanogen community.Item Animal factors that affect enteric methane production measured using the GreenFeed monitoring system in grazing dairy cows.(Elsevier B.V., 2024-04-16) Starsmore K; Lopez-Villalobos N; Shalloo L; Egan M; Burke J; Lahart BSimilar to all dairy systems internationally, pasture-based dairy systems are under increasing pressure to reduce their greenhouse gas (GHG) emissions. Ireland and New Zealand are 2 countries operating predominantly pasture-based dairy production systems where enteric CH4 contributes 23% and 36% of total national emissions, respectively. Ireland currently has a national commitment to reduce 51% of total GHG emissions by 2030 and 25% from agriculture by 2030, as well as striving to achieve climate neutrality by 2050. New Zealand's national commitment is to reduce 10% of methane emissions by 2030 and between 24% and 47% reduction in methane emissions by 2050. To achieve these reductions, factors that affect enteric methane (CH4) production in a pasture-based system need to be investigated. The objective of this study was to assess the relationship between enteric CH4 and other animal traits (feed intake, metabolic liveweight, energy corrected milk yield, milk urea concentration, and body condition score [BCS]) in a grazing dairy system. Enteric CH4 emissions were measured on 45 late lactation (213.8 ± 29 d after calving) grazing Holstein-Friesian and Holstein-Friesian × Jersey crossbred cows (lactation number 3.01 ± 1.65, 538.64 ± 59.37 kg live weight, and 3.14 ± 0.26 BCS) using GreenFeed monitoring equipment for 10 wk. There was a training period for the cows to use the GreenFeed of 3 wk before the 10-wk study period. The average enteric CH4 produced in the study was 352 g ± 45.7 g per day with an animal to animal coefficient of variation of 13%. Dry matter intake averaged 16.6 kg ± 2.23 kg per day, while milk solids (fat plus protein) averaged 1.62 kg ± 0.29 kg per day. A multiple linear regression model indicated that each one unit increase in energy corrected milk yield, metabolic liveweight and milk urea concentration, resulted in an increase in enteric CH4 production per day by 3.9, 1.74, and 1.38 g, respectively. Although each one unit increase in BCS resulted in a decrease in 39.03 g CH4 produced per day. When combined, these factors explained 47% of the variation in CH4 production, indicating that there is a large proportion of variation not included in the model. The repeatability of the CH4 measurements was 0.66 indicating that cows are relatively consistently exhibiting the same level of CH4 throughout the study. Therefore, enteric CH4 production is suitable for phenotyping.Item Effect of 3-nitrooxypropanol on enteric methane emissions of feedlot cattle fed with a tempered barley-based diet with canola oil(Oxford University Press on behalf of the American Society of Animal Science, 2023-07-10) Almeida AK; Cowley F; McMeniman JP; Karagiannis A; Walker N; Tamassia LFM; McGrath JJ; Hegarty RSA dose-response experiment was designed to examine the effect of 3-nitrooxypropanol (3-NOP) on methane (CH4) emissions, rumen function and performance of feedlot cattle fed a tempered barley-based diet with canola oil. Twenty Angus steers of initial body weight (BW) of 356 ± 14.4 kg were allocated in a randomized complete block design. Initial BW was used as the blocking criterion. Cattle were housed in individual indoor pens for 112 d, including the first 21 d of adaptation followed by a 90-d finishing period when five different 3-NOP inclusion rates were compared: 0 mg/kg dry matter (DM; control), 50 mg/kg DM, 75 mg/kg DM, 100 mg/kg DM, and 125 mg/kg DM. Daily CH4 production was measured on day 7 (last day of starter diet), day 14 (last day of the first intermediate diet), and day 21 (last day of the second intermediate diet) of the adaptation period and on days 28, 49, 70, 91, and 112 of the finisher period using open circuit respiration chambers. Rumen digesta samples were collected from each steer on the day prior to chamber measurement postfeeding, and prefeeding on the day after the chamber measurement, for determination of rumen volatile fatty acids (VFA), ammonium-N, protozoa enumeration, pH, and reduction potential. Dry matter intake (DMI) was recorded daily and BW weekly. Data were analyzed in a mixed model including period, 3-NOP dose and their interaction as fixed effects, and block as a random effect. Our results demonstrated both a linear and quadratic (decreasing rate of change) effect on CH4 production (g/d) and CH4 yield (g/kg DMI) as 3-NOP dose increased (P < 0.01). The achieved mitigation for CH4 yield in our study ranged from approximately 65.5% up to 87.6% relative to control steers fed a finishing feedlot diet. Our results revealed that 3-NOP dose did not alter rumen fermentation parameters such as ammonium-N, VFA concentration nor VFA molar proportions. Although this experimental design was not focused on the effect of 3-NOP dose on feedlot performance, no negative effects of any 3-NOP dose were detected on animal production parameters. Ultimately, the knowledge on the CH4 suppression pattern of 3-NOP may facilitate sustainable pathways for the feedlot industry to lower its carbon footprint.Item Bioactive metabolites of Asparagopsis stabilized in canola oil completely suppress methane emissions in beef cattle fed a feedlot diet(Oxford University Press on behalf of the American Society of Animal Science., 2024-04-22) Cowley FC; Kinley RD; Mackenzie SL; Fortes MRS; Palmieri C; Simanungkalit G; Almeida AK; Roque BMAsparagopsis taxiformis (Asparagopsis) has been shown to be highly efficacious at inhibiting the production of methane (CH4) in ruminants. To date, Asparagopsis has been primarily produced as a dietary supplement by freeze-drying to retain the volatile bioactive compound bromoform (CHBr3) in the product. Steeping of Asparagopsis bioactive compounds into a vegetable oil carrier (Asp-Oil) is an alternative method of stabilizing Asparagopsis as a ruminant feed additive. A dose-response experimental design used 3 Asp-Oil-canola oil blends, low, medium, and high Asp-Oil which provided 17, 34, and 51 mg Asparagopsis derived CHBr3/kg dry matter intake (DMI), respectively (in addition to a zero CHBr3 canola oil control), in a tempered-barley based feedlot finisher diet, fed for 59 d to 20 Angus heifers (five replicates per treatment). On four occasions, live weight was measured and CH4 emissions were quantified in respiration chambers, and blood, rumen fluid, and fecal samples were collected. At the end of the experiment, all animals were slaughtered, with carcasses graded, and samples of meat and edible offal collected for testing of consumer sensory qualities and residues of CHBr3, bromide, and iodide. All Asp-Oil treatments reduced CH4 yield (g CH4/kg DMI, P = 0.008) from control levels, with the low, medium, and high Asp-Oil achieving 64%, 98%, and 99% reduction, respectively. Dissolved hydrogen increased linearly with increasing Asp-Oil inclusion, by more than 17-fold in the high Asp-Oil group (P = 0.017). There was no effect of Asp-Oil treatment on rumen temperature, pH, reduction potential, volatile fatty acid and ammonia production, rumen pathology, and histopathology (P > 0.10). There were no differences in animal production and carcass parameters (P > 0.10). There was no detectable CHBr3 in feces or any carcass samples (P > 0.10), and iodide and bromide residues in kidneys were at levels unlikely to lead to consumers exceeding recommended maximum intakes. Overall, Asp-Oil was found to be safe for animals and consumers of meat, and effective at reducing CH4 emissions and yield by up to 99% within the range of inclusion levels tested.Item Genomic insights into the physiology of Quinella, an iconic uncultured rumen bacterium.(Nature Portfolio, 2022-10-20) Kumar S; Altermann E; Leahy SC; Jauregui R; Jonker A; Henderson G; Kittelmann S; Attwood GT; Kamke J; Waters SM; Patchett ML; Janssen PHQuinella is a genus of iconic rumen bacteria first reported in 1913. There are no cultures of these bacteria, and information on their physiology is scarce and contradictory. Increased abundance of Quinella was previously found in the rumens of some sheep that emit low amounts of methane (CH4) relative to their feed intake, but whether Quinella contributes to low CH4 emissions is not known. Here, we concentrate Quinella cells from sheep rumen contents, extract and sequence DNA, and reconstruct Quinella genomes that are >90% complete with as little as 0.20% contamination. Bioinformatic analyses of the encoded proteins indicate that lactate and propionate formation are major fermentation pathways. The presence of a gene encoding a potential uptake hydrogenase suggests that Quinella might be able to use free hydrogen (H2). None of the inferred metabolic pathways is predicted to produce H2, a major precursor of CH4, which is consistent with the lower CH4 emissions from those sheep with high abundances of this bacterium.Item Hydrogen cross-feeders of the human gastrointestinal tract.(Taylor & Francis Group, 2019-01-01) Smith NW; Shorten PR; Altermann EH; Roy NC; McNabb WCHydrogen plays a key role in many microbial metabolic pathways in the human gastrointestinal tract (GIT) that have an impact on human nutrition, health and wellbeing. Hydrogen is produced by many members of the GIT microbiota, and may be subsequently utilized by cross-feeding microbes for growth and in the production of larger molecules. Hydrogenotrophic microbes fall into three functional groups: sulfate-reducing bacteria, methanogenic archaea and acetogenic bacteria, which can convert hydrogen into hydrogen sulfide, methane and acetate, respectively. Despite different energy yields per molecule of hydrogen used between the functional groups, all three can coexist in the human GIT. The factors affecting the numerical balance of hydrogenotrophs in the GIT remain unconfirmed. There is increasing evidence linking both hydrogen sulfide and methane to GIT diseases such as irritable bowel syndrome, and strategies for the mitigation of such health problems through targeting of hydrogenotrophs constitute an important field for further investigation.Item Examination of hydrogen cross-feeders using a colonic microbiota model(BioMed Central Ltd, 2021-12) Smith NW; Shorten PR; Altermann E; Roy NC; McNabb WCBACKGROUND: Hydrogen cross-feeding microbes form a functionally important subset of the human colonic microbiota. The three major hydrogenotrophic functional groups of the colon: sulphate-reducing bacteria (SRB), methanogens and reductive acetogens, have been linked to wide ranging impacts on host physiology, health and wellbeing. RESULTS: An existing mathematical model for microbial community growth and metabolism was combined with models for each of the three hydrogenotrophic functional groups. The model was further developed for application to the colonic environment via inclusion of responsive pH, host metabolite absorption and the inclusion of host mucins. Predictions of the model, using two existing metabolic parameter sets, were compared to experimental faecal culture datasets. Model accuracy varied between experiments and measured variables and was most successful in predicting the growth of high relative abundance functional groups, such as the Bacteroides, and short chain fatty acid (SCFA) production. Two versions of the colonic model were developed: one representing the colon with sequential compartments and one utilising a continuous spatial representation. When applied to the colonic environment, the model predicted pH dynamics within the ranges measured in vivo and SCFA ratios comparable to those in the literature. The continuous version of the model simulated relative abundances of microbial functional groups comparable to measured values, but predictions were sensitive to the metabolic parameter values used for each functional group. Sulphate availability was found to strongly influence hydrogenotroph activity in the continuous version of the model, correlating positively with SRB and sulphide concentration and negatively with methanogen concentration, but had no effect in the compartmentalised model version. CONCLUSIONS: Although the model predictions compared well to only some experimental measurements, the important features of the colon environment included make it a novel and useful contribution to modelling the colonic microbiota.Item In vitro gas production and rumen fermentation profile of fresh and ensiled genetically modified high–metabolizable energy ryegrass(FASS Inc. and Elsevier Inc. on behalf of the American Dairy Science Association, 2020-03) Winichayakul S; Beechey-Gradwell Z; Muetzel S; Molano G; Crowther T; Lewis S; Xue H; Burke J; Bryan G; Roberts NJWe previously generated a high–metabolizable energy (HME) perennial ryegrass (Lolium perenne) by genetically modifying the plant to increase the leaf lipid content. Although substantial progress has been made toward characterizing physiological changes of HME ryegrass, very limited information exists for feeding value and its suitability for adoption into the pastoral system. In this study, independent HME ryegrass lines with a range of elevated leaf lipid concentrations were analyzed for changes in fatty acids and possible associated changes in the broader nutritional profile, including the gross energy, which was found to increase by 6.8%. Because ryegrass is often ensiled and fermentation in the rumen leads to biohydrogenation of fatty acids as well as enteric methane production, we sought to investigate these effects on HME ryegrass. This was achieved by performing mini-scale silos and using an automated gas measurement system to incubate the material in rumen fluid in vitro for 24 h. Our study included treatments comprising 3 independent HME ryegrass genotypes and wild-type control materials prepared fresh and as silage, employing in total 5 incubation studies, using rumen fluids collected from 4 nonlactating Jersey × Holstein cows. At intervals during the incubation, the production of gases, volatile fatty acids, and the degree of biohydrogenation were measured. Statistical data analysis indicated that differences in the nutritional compositions of the ensiled materials largely reflected those of their fresh counterparts. Incubation of both fresh and ensiled HME ryegrass in rumen fluid resulted in: (1) a greater percentage of valuable unsaturated fatty acids compared with the control; (2) a significant reduction of butyrate; and (3) a 10 to 15% decrease in the methane proportion of the total gas production. We conclude that ensiling could be a convenient option for preserving HME as a locally produced high-value supplementary feed; however, large-scale application needs to be investigated. In this paper we discuss the potential use of HME ryegrass to enhancing forage feeding value and the potential environmental benefits to the pastoral agriculture industry.
