Association of fibre degradation with ruminal dissolved hydrogen in growing beef bulls fed with two types of forages.

The present study investigated the association between fibre degradation and the concentration of dissolved molecular hydrogen (H2) in the rumen. Napier grass (NG) silage and corn stover (CS) silage were compared as forages with contrasting structures and degradation patterns. In the first experiment, CS silage had greater 48-h DM, neutral-detergent fibre (NDF) and acid-detergent fibre degradation, and total gas and methane (CH4) volumes, and lower 48-h H2 volume than NG silage in 48-h in vitro incubations. In the second experiment, twenty-four growing beef bulls were fed diets including 55 % (DM basis) NG or CS silages. Bulls fed the CS diet had greater DM intake (DMI), average daily gain, total-tract digestibility of OM and NDF, ruminal dissolved methane (dCH4) concentration and gene copies of protozoa, methanogens, Ruminococcus albus and R. flavefaciens, and had lower ruminal dH2 concentration, and molar proportions of valerate and isovalerate, in comparison with those fed the NG diet. There was a negative correlation between dH2 concentration and NDF digestibility in bulls fed the CS diet, and a lack of relationship between dH2 concentration and NDF digestibility with the NG diet. In summary, the fibre of CS silage was more easily degraded by rumen microorganisms than that of NG silage. Increased dCH4 concentration with the CS diet presumably led to the decreased ruminal dH2 concentration, which may be helpful for fibre degradation and growth of fibrolytic micro-organisms in the rumen.

Effects of rumen cannulation on dissolved gases and methanogen community in dairy cows.

Rumen cannulation is a widely employed technique in ruminant nutrition research. However, the gap between skin and rumen cannula can cause leakage of fermentation gases and influx of atmospheric air, which may adversely affect the anaerobic environment in the rumen. The present study was designed to investigate the effects of rumen cannulation on headspace gases, dissolved gases, fermentation end products, and methanogen community in the rumen of dairy cows. Eight Holstein cows were used in the experiment. Four cows were surgically fitted with rumen cannulas, whereas the other 4 intact cows were used as control. Rumen cannulation decreased gaseous hydrogen and methane concentrations, dissolved carbon dioxide concentration, and relative abundances of Methanosphaera, and increased the saturation factor of dissolved hydrogen and dissolved methane, dissolved methane concentration, volatile fatty acid concentration, 16S ribosomal RNA gene copies of methanogens, and Simpson index of methanogen community. In summary, rumen cannulation causes a reduction in headspace gaseous hydrogen and gaseous methane, which may not decrease dissolved gas concentrations due to an increase in saturation factors. Furthermore, rumen cannulation alters methanogen community with increased methanogen population and decreased relative abundances of Methanosphaera.

Technical note: Evaluation of interval between measurements and calculation method for the quantification of enteric methane emissions measured by respiration chamber.

Respiration chambers share one analyzer working in parallel, and methane (CH4) concentrations have to be measured at certain intervals. The maximum and minimum values in the kinetics of CH4 emissions can be missed during the interval between measurements, which may influence the quantification of CH4 emissions. Chambers must be opened for morning feeding and cleaning, which causes a loss of CH4 data. Calculation methods are needed to estimate the lost CH4 emission data, which may influence the estimated amount of daily CH4 emissions. In this study, we measured the CH4 emissions of 10 growing Chinese Holstein dairy heifers in respiration chambers. Methane concentrations were measured every 0.5 min to obtain the 23-h kinetics of CH4 emissions, which were further selected at different intervals between measurements (i.e., 5, 30, 60, 120, 180, and 240 min) to evaluate the effects of interval on quantification of CH4 emissions. The missing 1-h kinetics of CH4 emissions before feeding were not measured, and 2 calculation methods were used to estimate the missing 1-h kinetics of CH4 emissions: mean value of measuring period (the mean method) and the nearest value of measurement just before chamber opening (the nearest method). The results showed that the rates of CH4 emission from 10 heifers varied from 4.56 to 11.42 g/h. The increment of intervals decreased maximum rate of CH4 emission and increased minimum rate of CH4 emission. Interval caused less than 5% of the difference in measuring CH4 emissions. Although the mean method had greater estimated daily CH4 emission than the nearest method, the difference was within 3%. The interval between measurements (≤3 h) and calculation method had little influence on enteric CH4 emission measurements.

Nitrate improves ammonia incorporation into rumen microbial protein in lactating dairy cows fed a low-protein diet.

Generation of ammonia from nitrate reduction is slower compared with urea hydrolysis and may be more efficiently incorporated into ruminal microbial protein. We hypothesized that nitrate supplementation could increase ammonia incorporation into microbial protein in the rumen compared with urea supplementation of a low-protein diet fed to lactating dairy cows. Eight multiparous Chinese Holstein dairy cows were used in a crossover design to investigate the effect of nitrate or an isonitrogenous urea inclusion in the basal low-protein diet on rumen fermentation, milk yield, and ruminal microbial community in dairy cows fed a low-protein diet in comparison with an isonitrogenous urea control. Eight lactating cows were blocked in 4 pairs according to days in milk, parity, and milk yield and allocated to urea (7.0 g urea/kg of dry matter of basal diet) or nitrate (14.6 g of NO3-/kg of dry matter of basal diet, supplemented as sodium nitrate) treatments, which were formulated on 75% of metabolizable protein requirements. Nitrate supplementation decreased ammonia concentration in the rumen liquids (-33.1%) and plasma (-30.6%) as well as methane emissions (-15.0%) and increased dissolved hydrogen concentration (102%), microbial N (22.8%), propionate molar percentage, milk yield, and 16S rRNA gene copies of Selenomonas ruminantium. Ruminal dissolved hydrogen was positively correlated with the molar proportion of propionate (r = 0.57), and negatively correlated with acetate-to-propionate ratio (r = -0.57) and estimated net metabolic hydrogen production relative to total VFA produced (r = -0.58). Nitrate reduction to ammonia redirected metabolic hydrogen away from methanogenesis, enhanced ammonia incorporation into rumen microbial protein, and shifted fermentation from acetate to propionate, along with increasing S. ruminantium 16S rRNA gene copies, likely leading to the increased milk yield.

Effects of three methane mitigation agents on parameters of kinetics of total and hydrogen gas production, ruminal fermentation and hydrogen balance using in vitro technique.

Methane (CH4 ) can be mitigated through directly inhibiting methanogen activity and starving methanogens by hydrogen (H2 ) sink. Three types of mechanism (i.e. bromoethanesulphonate (BES), nitrate and emodin) and doses of CH4 mitigation agents were employed to investigate their pathways of CH4 inhibition. Results indicated that both BES and emodin inhibited CH4 production and altered H2 balance, which could be accompanied by decreased dry matter disappearance (DMD), fractional rate of gH2 formation, volatile fatty acid (VFA) production, ability to produce and use reducing equivalences and molecular H2 , and increased final asymptotic gH2 production, time to the peak of gH2 , discrete lag time of gH2 production and fermentation efficiency. However, emodin decreased gas volume produced by rapidly fermentable components of substrate and the rate of fermentation at early stage of incubation, while BES supplementation inhibited gas volume produced by both rapidly and slowly fermentable components of substrate and the rate of fermentation at middle or late stage of incubation. The nitrate supplementation inhibited CH4 production without affecting VFA profile, because of its dual role as H2 sink and being toxic to methanogens. Nitrate supplementation had more complicated pattern of fermentation, VFA production and profile and H2 balance in comparison to BES and emodin supplementation.

[Accuracy of predicting in vitro ruminal methane production in goats using volatile fatty acids stoichiometric models].

This study was conducted to investigate the accuracy of predicting in vitro ruminal methane (CH4) production using volatile fatty acids (VFA) stoichiometric models [CH4 = 0.5Ace-0.25Pro + 0.5But-0.25Val] (model 1), where CH4, Ace, Pro, But and Val are the production amounts of CH4, acetate, propionate, butyrate and valerate, respectively. Ten common feedstuffs, including four concentrates and six roughages with a wide range of chemical composition were incubated in serum bottles, and VFAs and CH4 production at 72 h were determined. The differences between the predicted and measured CH4 production were quantified using the model accuracy analysis. The results showed that the predicted CH4 production amounts were generally greater than the measured values obtained using the model 1, and the bias, slope and random error were 62.6%, 11.7% and 25.7%, respectively, indicating that fixed error exceeded 70%. By assuming 80% of total hydrogen being used for CH4 synthesis, the VFA stoichiometric model could be re-expressed as [CH4 = 0.8 (0.5Ace-0.25Pro + 0.5But-0.25Val)] (model 2). The root mean square prediction error (rMSPE = 0.18) for model 2 was less than for model 1 (rMSPE = 0.60). In addition, the bias, slope and random error of the model 2 were 2.1%, 5.7%, 92.3%, respectively, indicating that fixed error was less than 10%. In model 1, hydrogen formation resulting from VFA production were assumed to be totally consumed by methanogens for CH4 synthesis, without considering other pathways of hydrogen metabolism, which was the main factor resulting in the higher predicted values than the measured values.