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Nitroethane (NE), 2-nitroethanol (NEOH) and 2-nitro-1-propanol (NPOH) were investigated in order to determine their inhibitory effects on in vitro ruminal fermentation and methane (CH4) production of a hay-rich substrate (alfalfa hay: maize meal = 4:1, w/w). The rumen liquor collected from cannulated Holstein dairy cows was incubated at 39 °C for 72 h. The addition of NE, NEOH and NPOH slowed down the fermentation process and notably decreased molar CH4 proportion by 96.8, 96.4 and 35.0%, respectively. The abundance of total methanogen and methanogens from the order Methanobacteriales were all decreased with NE, NEOH and NPOH supplementation. Meanwhile, the nitrocompound addition reduced mcrA gene expression, coenzyme F420 and F430 contents. The correlation analysis showed that CH4 production was correlated positively with the population abundance of total methanogens, Methanobacteriales, mcrA gene expression, coenzyme contents of F420 and F430. The nitrocompound addition decreased acetate concentration and increased propionate and butyrate concentrations in the culture fluid. In summary, both NE and NEOH addition presented nearly the same inhibitory effectiveness on in vitro CH4 production; they were more effective than NPOH. The results of the current study provide evidence that NE, NEOH and NPOH can dramatically decrease methanogen population, mcrA gene expression and the coenzyme content of F420 and F430 in ruminal methanogenesis.

In vivo and in vitro trials were conducted to assess the effects of tributyrin (TB) supplementation on short-chain fatty acid (SFCA) concentrations, fibrolytic enzyme activity, nutrient digestibility and methanogenesis in adult sheep. Nine 12-month-old ruminally cannulated Small Tail ewes (initial body weight 55 ± 5.0 kg) without pregnancy were used for the in vitro trial. In vitro substrate made to offer TB at 0, 2, 4, 6 and 8 g/kg on a dry matter (DM) basis was incubated by ruminal microbes for 72 h at 39°C. Forty-five adult Small Tail ewes used for the in vivo trial were randomly assigned to five treatments with nine animals each for an 18-d period according to body weight (55 ± 5.0 kg). Total mixed ration fed to ewes was also used to offer TB at 0, 2, 4, 6 and 8 g/kg on a DM basis. The in vitro trial showed that TB supplementation linearly increased apparent digestibility of DM, crude protein, neutral detergent fibre and acid detergent fibre, and enhanced gas production and methane emissions. The in vivo trial showed that TB supplementation decreased DM intake, but enhanced ruminal fermentation efficiency. Both in vitro and in vivo trials showed that TB supplementation enhanced total SFCA concentrations and carboxymethyl cellulase activity. The results indicate that TB supplementation might exert advantage effects on rumen microbial metabolism, despite having an enhancing effect on methanogenesis.

SUMMARYThe present study investigated the effects of different levels of urea nitrogen (N) fertilizer on nutrient accumulation,in vitrorumen gas production and fermentation characteristics of forage oat straw (FOS) from oats (Avena sativaL. ‘Qinghai 444’) grown in the Tibet region of China. Fertilizer, applied at seeding (day 1), stem elongation (days 52–54) and heading (days 63–67), increased plant height and prolonged the maturity stage of the plant by 4–11 days compared with the non-fertilized control. Oat plants were harvested at maturity at the node 3–4 cm above ground, and then separated into grains and FOS. Both FOS and grain yields increased quadratically with increasing N fertilization, and their theoretical maximums occurred at the N fertilizing rates of 439 and 385 kg/ha, respectively. Increases in N fertilization did not affect the hemicellulose content of FOS, but substantially promoted the accumulation of crude protein, cellulose and lignin, resulting in a decrease in the energy content available for metabolism. A 72-h incubation of FOS with rumen fluids from lactating cows showed that increasing N resulted in FOS that showed a slower fermentation rate, decreasedin vitrodry matter disappearance and lower cumulative gas production, but unchanged fermentation gas composition. Nitrogen fertilization increased the final pH in culture fluids and decreased the microbial volatile fatty acid (VFA) production. The molar proportions of acetate and propionate were not affected, but molar propionate proportion decreased linearly with increasing urea fertilization, and consequently, the ratio of lipogenic (e.g., acetate and butyrate)-to-glucogenic acids (propionate) tended to increase. In brief, increasing urea N fertilization promoted the growth of forage oats and increased the biomass yield as well as the crude protein and cellulose content of FOS. Considering the negative effect of increased lignin content on nutrient digestibility and total VFA production, the suggested range of urea N fertilization is 156–363 kg N/ha for forage oats planted in Tibet to retain the nutritive value of FOS in the rumen.

SUMMARYA completely randomized experiment for planting highland barley in 36 field plots of the Lhasa Agricultural Experiment Station was applied to investigate the effect of urea nitrogen (N) fertilization levels of 0 (control), 156, 258, 363, 465 and 570 kg/ha on nutrient accumulation,in vitrorumen gas production and fermentation characteristics of highland barley straw (HBS). Each urea application was divided into three portions of 0.4, 0.3 and 0.3 and sequentially fertilized at seeding (growth stage (GS) 0), stem elongation (GS 32) and heading (GS 49), respectively. The maturity stage lasted 5–13 days longer in response to the urea N fertilization compared with the control. After removing grains, HBS biomass was harvested at maturity. The biomass yields of leaf, stem, straw and grain were increased quadratically with increasing urea N fertilization, and HBS and grain yields peaked at the estimated urea N fertilization levels of 385 and 428 kg/ha, respectively. The increase of urea N fertilization increased the accumulation of crude protein, cellulose and lignin, and decreased the content of ash and hemicellulose in HBS, resulting in a decrease of the energy content available to be metabolized. After incubating HBS for 72 h with rumen fluids from lactating cows, the urea N fertilization decreasedin vitrodry matter disappearance and cumulative gas production, and slightly altered fermentation end-gas composition. Urea N fertilization decreased microbial volatile fatty acid production, but did not alter the ratio of lipogenic acetate and butyrate to glucogenic propionate. In a brief, the current urea N fertilization strategy promoted the growth of the highland barley and increased biomass yield, protein and cellulose accumulation of HBS. A urea N fertilization level ⩽385 kg/ha could be sufficient for growth of highland barley in Tibet without a consequent nutritive reduction in ruminal digestion.

SUMMARYLignification-associated phenolic acids are widely distributed in graminaceous plant cell walls. Nylon bags containing maize bran, wheat bran, millet husk and rice husk were incubated in the rumens of five Charolais (♂) × Nanyang (♀) crossbred steers for 6, 12, 24, 36, 48 and 72 h. The in situ ruminal disappearance of ester-linked phenolic acids linearly increased in the brans with increasing incubation time, and the disappearance was greater for ester-linked ferulic acid (FAest) than for ester-linked p-coumaric acid (PCAest). The disappearances of FAest and PCAest were positively correlated with disappearances of neutral detergent fibre (NDF), cellulose and hemicellulose. The effective degradabilities of NDF, cellulose and hemicellulose in the brans were markedly greater than the effective degradabilities of these components in the husks, and were negatively correlated with the contents of Lignin (sa), ether-linked ferulic acid, PCAest and ether-linked p-coumaric acid in both the cereal brans and husks. These findings suggested that breeding forage crops with modified phenolic acid contents could represent an alternative strategy to promote further increases in fibre digestibility of cereal residue feeds for ruminant animals.

SUMMARYThe objective of the present study was to investigate the effect of dietary neutral detergent fibre (NDF) : crude protein (CP) ratio on duodenal microbial crude protein (MCP) flow and nitrogen (N) losses. The study was completed in a 5 × 5 Latin square design with five lactating Holstein dairy cows and 5 high-concentrate total mixed rations (TMR) with different forage combinations, typical for Northern China. The rations with a fixed forage-to-concentrate ratio (39 : 61) resulted in different dietary NDF : CP ratios: TMR1 3·03 : 1 (428·2 g NDF/kg : 141·4 g CP/kg); TMR2 2·74 : 1 (392·7 g NDF/kg : 143·2 g CP/kg); TMR3 2·55 : 1 (368·3 g NDF/kg : 144·4 g CP/kg); TMR4 1·84 : 1 (304·8 g NDF/kg : 165·8 g CP/kg); TMR5 1·60 : 1 (285·0 g NDF/kg : 178·0 g CP/kg). Rumen content, milk, blood, urine and faeces were sampled on the last 3 days of five 18-day periods. Purine derivatives in the urine samples were determined to estimate rumen MCP flow into the small intestine. Milk yield and milk protein yield increased linearly with decreasing dietary NDF : CP ratio although slight differences in dry matter intake were observed due to feed intake restriction. Diurnal ammonia N in the rumen and duodenal MCP flow increased linearly, but blood urea N, urinary N and faecal N linearly decreased with decreasing dietary NDF : CP ratio. The enhanced N utilization in the maize-silage-based TMRs (TMR4–5) in comparison with maize-stover-based TMRs (TMR1–3) increased milk yield and the synthesis of milk protein instead of milk fat in the lactating cows, probably due to high transfer of ammonia N into rumen MCP with a considerable increase of dietary non-fibre carbohydrate content and the decrease of NDF : CP ratio. The present results indicate that not only increasing dietary non-structural carbohydrate content but also adjusting the ratio of structural carbohydrate to CP ratio are important diet formulation strategies for mitigating N losses in lactating cows.