Anthony Waterhouse

Bovine Host Genetic Variation Influences Rumen Microbial Methane Production with Best Selection Criterion for Low Methane Emitting and Efficiently Feed Converting Hosts Based on Metagenomic Gene Abundance.

Methane produced by methanogenic archaea in ruminants contributes significantly to anthropogenic greenhouse gas emissions. The host genetic link controlling microbial methane production is unknown and appropriate genetic selection strategies are not developed. We used sire progeny group differences to estimate the host genetic influence on rumen microbial methane production in a factorial experiment consisting of crossbred breed types and diets. Rumen metagenomic profiling was undertaken to investigate links between microbial genes and methane emissions or feed conversion efficiency. Sire progeny groups differed significantly in their methane emissions measured in respiration chambers. Ranking of the sire progeny groups based on methane emissions or relative archaeal abundance was consistent overall and within diet, suggesting that archaeal abundance in ruminal digesta is under host genetic control and can be used to genetically select animals without measuring methane directly. In the metagenomic analysis of rumen contents, we identified 3970 microbial genes of which 20 and 49 genes were significantly associated with methane emissions and feed conversion efficiency respectively. These explained 81% and 86% of the respective variation and were clustered in distinct functional gene networks. Methanogenesis genes (e.g. mcrA and fmdB) were associated with methane emissions, whilst host-microbiome cross talk genes (e.g. TSTA3 and FucI) were associated with feed conversion efficiency. These results strengthen the idea that the host animal controls its own microbiota to a significant extent and open up the implementation of effective breeding strategies using rumen microbial gene abundance as a predictor for difficult-to-measure traits on a large number of hosts. Generally, the results provide a proof of principle to use the relative abundance of microbial genes in the gastrointestinal tract of different species to predict their influence on traits e.g. human metabolism, health and behaviour, as well as to understand the genetic link between host and microbiome.

Archaeal abundance in post-mortem ruminal digesta may help predict methane emissions from beef cattle

Methane produced from 35 Aberdeen-Angus and 33 Limousin cross steers was measured in respiration chambers. Each group was split to receive either a medium- or high-concentrate diet. Ruminal digesta samples were subsequently removed to investigate correlations between methane emissions and the rumen microbial community, as measured by qPCR of 16S or 18S rRNA genes. Diet had the greatest influence on methane emissions. The high-concentrate diet resulted in lower methane emissions (P < 0.001) than the medium-concentrate diet. Methane was correlated, irrespective of breed, with the abundance of archaea (R = 0.39), bacteria (−0.47), protozoa (0.45), Bacteroidetes (−0.37) and Clostridium Cluster XIVa (−0.35). The archaea:bacteria ratio provided a stronger correlation (0.49). A similar correlation was found with digesta samples taken 2–3 weeks later at slaughter. This finding could help enable greenhouse gas emissions of large animal cohorts to be predicted from samples taken conveniently in the abattoir.

Effectiveness of nitrate addition and increased oil content as methane mitigation strategies for beef cattle fed two contrasting basal diets

The objectives of this study were to investigate the effects of (1) the addition of nitrate and (2) an increase in dietary oil on methane (CH4) and hydrogen (H2) emissions from 2 breeds (cross-bred Charolais and purebred Luing) of finishing beef cattle receiving 2 contrasting basal diets consisting (grams per kilogram DM) of 500:500 (Mixed) and 80:920 (Concentrate) forage to concentrate ratios. Within each basal diet there were 3 treatments: (i) control treatments (mixed-CTL and concentrate-CTL) contained rapeseed meal as the protein source, which was replaced with either (ii) calcium nitrate (mixed-NIT and concentrate-NIT) supplying 21.5 g nitrate/kg DM, or (iii) rapeseed cake (mixed-RSC and concentrate-RSC) to increase dietary oil from 27 (CTL) to 53 g/kg DM (RSC). Following adaption to diets, CH4 and H2 emissions were measured on 1 occasion from each of the 76 steers over a 13-wk period. Dry matter intakes tended (P = 0.051) to be greater for the concentrate diet than the mixed diet; however, when expressed as grams DMI per kilogram BW, there was no difference between diets (P = 0.41). Dry matter intakes for NIT or RSC did not differ from CTL. Steers fed a concentrate diet produced less CH4 and H2 than those fed a mixed diet (P < 0.001). Molar proportions of acetate (P < 0.001) and butyrate (P < 0.01) were lower and propionate (P < 0.001) and valerate (P < 0.05) higher in the rumen fluid from steers fed the concentrate diet. For the mixed diet, CH4 yield (grams per kilogram DMI) was decreased by 17% when nitrate was added (P < 0.01), while H2 yield increased by 160% (P < 0.001). The addition of RSC to the mixed diet decreased CH4 yield by 7.5% (P = 0.18). However, for the concentrate diet neither addition of nitrate (P = 0.65) nor increasing dietary oil content (P = 0.46) decreased CH4 yield compared to concentrate-CTL. Molar proportions of acetate were higher (P < 0.001) and those of propionate lower (P < 0.01) in rumen fluid from NIT treatments compared to respective CTL treatments. Overall, reductions in CH4 emissions from adding nitrate or increasing the oil content of the mixed diet were similar to those expected from previous reports. However, the lack of an effect of these mitigation strategies when used with high concentrate diets has not been previously reported. This study shows that the effect of CH4 mitigation strategies is basal diet-dependent.

Evaluation of the laser methane detector to estimate methane emissions from ewes and steers.

The laser methane detector (LMD) has been proposed as a method to characterize enteric methane (CH4) emissions from animals in a natural environment. To validate LMD use, its CH4 outputs (LMD-CH4), were compared against CH4 measured with respiration chambers (chamber-CH4). The LMD was used to measure CH4 concentration (µL/L) in the exhaled air of 24 lactating ewes and 72 finishing steers. In ewes, LMD was used on 1 d for each ewe, for 2-min periods at 5 hourly observation periods (P1 to P5, respectively) after feeding. In steers fed either low- or high-concentrate diets, LMD was used once daily for a 4-min period for 3 d. The week after LMD-CH4 measurement, ewes or steers entered respiration chambers to quantify daily CH4 output (g/d). The LMD outputs consisted of periodic events of high CH4 concentrations superimposed on a background of oscillating lower CH4 concentrations. The high CH4 events were attributed to eructation and the lower background CH4 to respiration. After fitting a double normal distribution to the data set, a threshold of 99% of probability of the lower distribution was used to separate respiration from eructation events. The correlation between mean LMD-CH4 and chamber-CH4 was not high, and only improved correlations were observed after data were separated in 2 levels. In ewes, a model with LMD and DMI (adjusted R(2) = 0.92) improved the relationship between DMI and chamber-CH4 alone (adjusted R(2) = 0.79) and between LMD and chamber-CH4 alone (adjusted R(2) = 0.86). In both experiments, chamber-CH4 was best explained by models with length of eructation events (time) and maximum values of CH4 concentration during respiration events (µL/L; P < 0.01). Correlation between methods differed between observation periods, indicating the best results of the LMD were observed from 3 to 5 h after feeding. Given the short time and ease of use of LMD, there is potential for its commercial application and field-based studies. Although good indicators of quantity of CH4 were obtained with respiration and eructation CH4, the method needed to separate the data into high and low levels of CH4 was not simple to apply in practice. Further assessment of the LMD should be performed in relation to animal feeding behavior and physiology to validate assumptions of eructation and respiration levels, and other sources of variation should be tested (i.e., micrometeorology) to better investigate its potential application for CH4 testing in outdoor conditions.

Impact of adding nitrate or increasing the lipid content of two contrasting diets on blood methaemoglobin and performance of two breeds of finishing beef steers.

Adding nitrate to the diet or increasing the concentration of dietary lipid are effective strategies for reducing enteric methane emissions. This study investigated their effect on health and performance of finishing beef cattle. The experiment was a two×two×three factorial design comprising two breeds (CHX, crossbred Charolais; LU, Luing); two basal diets consisting of (g/kg dry matter (DM), forage to concentrate ratios) 520 : 480 (Mixed) or 84 : 916 (Concentrate); and three treatments: (i) control with rapeseed meal as the main protein source replaced with either (ii) calcium nitrate (18 g nitrate/kg diet DM) or (iii) rapeseed cake (RSC, increasing acid hydrolysed ether extract from 25 to 48 g/kg diet DM). Steers (n=84) were allocated to each of the six basal diet×treatments in equal numbers of each breed with feed offered ad libitum. Blood methaemoglobin (MetHb) concentrations (marker for nitrate poisoning) were monitored throughout the study in steers receiving nitrate. After dietary adaptation over 28 days, individual animal intake, performance and feed efficiency were recorded for a test period of 56 days. Blood MetHb concentrations were low and similar up to 14 g nitrate/kg diet DM but increased when nitrate increased to 18 g nitrate/kg diet DM (P0.05). Neither basal diet nor treatment affected carcass quality (P>0.05), but CHX steers achieved a greater killing out proportion (P<0.001) than LU steers. Thus, adding nitrate to the diet or increasing the level of dietary lipid through the use of cold-pressed RSC, did not adversely affect health or performance of finishing beef steers when used within the diets studied.

Does diverse grazing behavior of suckler cows have an impact on predicted methane emissions? 1

A modeling study based on a dataset from a large-scale grazing study was used to identify the potential impact of grazing behavior and performance of diverse cow genotypes on predicted methane (CH4) emissions. Lactating cows grazing extensive seminatural grassland and heath vegetation were monitored with Global Positioning System collars and activity sensors. The diet selected by cows of 3 different genotypes, Aberdeen Angus cross Limousin (AxL), Charolais (CHA), and Luing (LUI), was simulated by matching their locations during active periods with hill vegetation maps. Measured performance and activity were used to predict energy requirements, DMI, and CH4 output. The cumulative effect of actual performance, diet selection, and actual physical activity on potential CH4 output and yield was estimated. Sensitivity analyses were performed for the digestibility of intake, energy cost of activity, proportion of milk consumed by calves, and reproductive efficiency. Although with a better performance (P < 0.05), LUI required less total energy than the other genotypes (P < 0.001) as the other 2 spent more energy for maintenance (P < 0.001) and activity (P < 0.001). By selecting a better quality diet (P < 0.03), estimated CH4 of CHA cow-calf pairs was lower than AxL (P = 0.001) and slightly lower than LUI (P = 0.08). Energy lost as CH4 was 0.17 and 0.58% lower for LUI than AxL and CHA (P < 0.002). This study suggests for the first time that measured activity has a major impact on estimated CH4 outputs. A 15% difference of the cow-calf pair CH4 was estimated when using different coefficients to convert actual activity into energy. Predicted CH4 was highly sensitive to small changes in diet quality, suggesting the relative importance of diet selection on heterogeneous rangelands. Extending these results to a farm systems scale, CH4 outputs were also highly sensitive to reductions in weaning rates, illustrating the impact on CH4 at the farm-system level of using poorly adapted genotypes on habitats where their performances may be compromised. This paper demonstrates that variations in grazing behavior and grazing choice have a potentially large impact on CH4 emissions, illustrating the importance of including these factors in calculating realistic national and global estimates.

The impact of divergent breed types and diets on methane emissions, rumen characteristics and performance of finishing beef cattle

This study was undertaken to further develop our understanding of the links between breed, diet and the rumen microbial community and determine their effect on production characteristics and methane (CH4) emissions from beef cattle. The experiment was of a 2×2 factorial design, comprising two breeds (crossbred Charolais (CHX); purebred Luing (LU)) and two diets (concentrate-straw or silage-based). In total, 80 steers were used and balanced for sire within each breed, farm of origin and BW across diets. The diets (fed as total mixed rations) consisted of (g/kg dry matter (DM)) forage to concentrate ratios of either 500 : 500 (Mixed) or 79 : 921 (Concentrate). Steers were adapted to the diets over a 4-week period and performance and feed efficiency were then measured over a 56-day test period. Directly after the 56-day test, CH4 and carbon dioxide (CO2) emissions were measured (six steers/week) over a 13-week period. Compared with LU steers, CHX steers had greater average daily gain (ADG; P<0.05) and significantly (P<0.001) lower residual feed intake. Crossbred Charolais steers had superior conformation and fatness scores (P<0.001) than LU steers. Although steers consumed, on a DM basis, more Concentrate than Mixed diet (P<0.01), there were no differences between diets in either ADG or feed efficiency during the 56-day test. At slaughter, however, Concentrate-fed steers were heavier (P<0.05) and had greater carcass weights than Mixed-fed steers (P<0.001). Breed of steer did not influence CH4 production, but it was substantially lower when the Concentrate rather than Mixed diet was fed (P<0.001). Rumen fluid from Concentrate-fed steers contained greater proportions of propionic acid (P<0.001) and lower proportions of acetic acid (P<0.001), fewer archaea (P<0.01) and protozoa (P=0.09), but more Clostridium Cluster XIVa (P<0.01) and Bacteroides plus Prevotella (P<0.001) than Mixed-fed steers. When the CH4 to CO2 molar ratio was considered as a proxy method for CH4 production (g/kg DM intake), only weak relationships were found within diets. In conclusion, although feeding Concentrate and Mixed diets produced substantial differences in CH4 emissions and rumen characteristics, differences in performance were influenced more markedly by breed.