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Aims Ferulic acid esterase (FAE)-producing Lactobacillus species isolated from ensiled Elymus nutans growing on the Qinghai-Tibetan plateau were characterized, and effects of their application to the alfalfa ensiling process and the evidence to synergic effect between cellulase and FAE were investigated. Methods and results The results of 16S rRNA gene sequence and species-specific polymerase chain reaction amplification showed that two screened strains with high FAE activity were Lactobacillus plantarum A1 (LP) and L. brevis A3 (LBr). The optimum temperature and pH for the LP and LBr was 37°C and 6·4 respectively. The FAE exhibited a good stability at temperatures between 25 and 50°C and at pH values of 5·0-7·0. The two strains and a commercial cellulase (CE) were applied as additives to alfalfa silage. After 60 days of ensiling, the lactic acid in the control and CE groups were significantly lower than those of the other treatment groups. The neutral detergent fibre and acid detergent fibre contents in the LP group were significantly lower than those observed in the other groups. At the same time, the combination of CE and FAE-producing lactic acid bacteria synergistically improved the fermentation quality of the silage. Conclusions The addition of the FAE-producing strain of L. plantarum A1 to alfalfa silage improved its fermentation quality, and reduced the fibre content of the silage. Significance and impact of the study The screened homo-fermentative and FAE-producing strain of L. plantarum A1 could be a candidate strain in improving fermentation quality and fibre digestibility of ensiled forages.

Aims Characteristics of a strain Pediococcus pentosaceus Q6 isolated from Elymus nutans growing on the Tibetan plateau and its effects on E. nutans silage fermentation stored at low temperature were investigated. Methods and results Sugar fermentation pattern and growth profiles of the strain Q6 and its reference strain APP were characterized. The strain Q6 and APP were inoculated to E. nutans at ensiling respectively; and ensiled at different temperatures (10, 15 and 25°C) for 30, 60 and 90 days. The results indicated that Q6 could grow at pH 3·0 and at 4°C. In contrast to APP, Q6 could ferment mannitol, saccharose, sorbitol and rhamnose. Lower pH in Q6-treated silages fermented for 60 days at 10 and 15°C was found compared with the control and APP-treated groups. For the silages that were stored at 10 or 15°C, the greatest lactic acid content were detected in Q6-inoculated silages ensiled for 30 and 60 days respectively. There were no differences in pH and lactic acid content between Q6- and APP-treated silages ensiled at 10 and 15°C for 90 days respectively. Conclusions Inoculation of the strain P. pentosaceus Q6 could improve fermentation quality of ensiled E. nutans at the early stage of ensiling stored at low temperature (10 or 15°C). Significance and impact of the study The selection of P. pentosaceus inoculants for improving silage quality at low temperature, which provides a candidate strain to make high-quality silage in regions with frigid climate.

Under traditional management on the Qinghai-Tibetan Plateau, yaks () graze only on natural pasture without supplements and are forced to cope with sparse forage of low N content, especially in winter. In contrast, indigenous Tibetan yellow cattle () require supplements during the cold season. We hypothesized that, in response to harsh conditions, yaks cope with low N intakes better than cattle. To test this hypothesis, a study of whole-body N retention and urea kinetics was conducted in 2 concurrent 4 × 4 Latin squares, with 1 square using yaks and 1 square using cattle. Four isocaloric forage-concentrate diets differing in N concentrations (10.3, 19.5, 28.5, and 37.6 g N/kg DM) were formulated, and by design, DMI were similar between species and across diets. Urea kinetics were determined with continuous intravenous infusion of NN urea for 104 h, and total urine and feces were concomitantly collected. Urea production, urea recycling to the gut, and ruminal microbial protein synthesis all linearly increased ( < 0.001) with increasing dietary N in both yaks and cattle. Urinary N excretion was less ( = 0.04) and N retention was greater ( = 0.01) in yaks than in cattle. Urea production was greater in yaks than in cattle at the 3 lowest N diets but greater in cattle than in yaks at the highest N diet (species × diet, < 0.02). Urea N recycled to the gut ( < 0.001), recycled urea N captured by ruminal bacteria ( < 0.001), and ruminal microbial protein production ( = 0.05) were greater in yaks than in cattle. No more than 12% of urea recycling was through saliva, with no difference between species ( = 0.61). Glomerular filtration rate was lower ( = 0.05) in yaks than in cattle. The higher urea recycling and greater capture of recycled urea by ruminal microbes in yaks than in cattle suggest that yaks use mechanisms to utilize dietary N more efficiently than cattle, which may partially explain the better survival of yaks than cattle when fed low-N diets.

To study metabolic adaptation to harsh foraging conditions, an experiment was conducted to characterize and quantify N utilization efficiency and urea metabolism in Tibetan and fine-wool sheep fed 4 levels of dietary N (11.0, 16.7, 23.1, and 29.2 g N/kg DM) in 2 concurrent 4 × 4 Latin square designs. Urea kinetics were determined using continuous intrajugular infusions of 15N15N-urea. Urinary excretions of total N and urea N increased linearly (P < 0.001) with dietary N and were not different between breeds (P ≥ 0.37). Fecal N excretion increased with dietary N for Tibetan sheep but not for fine-wool sheep (linear dietary N × breed; P < 0.05). Nitrogen retention (both amount per day and percentage of N intake) increased with increasing dietary N concentration (P < 0.001), and the rates of increase were greater in fine-wool than in Tibetan sheep (linear dietary N × breed and cubic dietary N × breed; P < 0.05). In Tibetan sheep, N retention as a percentage of intake was greatest for diets containing 16.7 g N/kg DM, whereas it was maximal for fine-wool sheep when the diet contained 23.1 g N/kg DM. Urea N entry rate, urea N recycled to the gastrointestinal tract (GIT), and urea N returned to the ornithine cycle all increased with dietary N (P < 0.05), and all were greater in Tibetan than fine-wool sheep for the 11.0 g N/kg DM diet but were greater in fine-wool than Tibetan sheep for the diet with 29.2 g N/kg DM (linear dietary N × breed; P < 0.05). Urea N excreted in feces, both amount and fraction of GIT entry rate, was less in Tibetan than fine-wool sheep for the 11.0 and 16.7 g N/kg DM diets but similar for diets with 23.1 or 29.2 g N/kg DM (linear dietary N × breed; P < 0.01). For the lowest-protein diet, the fraction of urea N production recycled to the GIT was greater in the Tibetan than fine-wool sheep (88% vs. 82%), but for the diet with 29.2 g N/kg DM it was greater for fine-wool than Tibetan sheep (46% vs. 39%; linear dietary N × breed; P < 0.05). Plasma urea N increased more rapidly in response to increasing dietary N concentration for fine-wool sheep than for Tibetan sheep (linear dietary N × breed; P < 0.05). Urea tubular load and the amount and percentage of urea reabsorbed by the kidney were greater in Tibetan than fine-wool sheep (P < 0.05). These results suggest that Tibetan sheep have mechanisms that allow them to utilize N more efficiently than the fine-wool sheep when dietary N is inadequate.

Two experiments were conducted to determine the urinary purine derivative (PD) excretion rates and to analyze potential variations in PD excretions between yak, Tibetan indigenous cattle, and their crossbreds under similar ecological environment, and to develop prediction equations based on PD excretion for estimating rumen microbial protein supply. Three Tibetan castrated male yaks, and similar numbers of indigenous cattle and their crossbreds, were used in a fasting trial followed by a feeding trial, using three 3 x 3 Latin squares (1 for each breedtype), to measure responses of PD excretion to different feeding levels. The results showed that i) daily endogenous PD excretion for yak, indigenous cattle, and their crossbreds was 134, 163, and 138 micromol/kg of BW(0.75) (P = 0.38), respectively; and ii) crossbreds have greater PD excretion rate per unit digestible OM intake than indigenous cattle (P = 0.03). However, there were no significant differences between yaks and the crossbreds (P = 0.24) or between yaks and the indigenous cattle (P = 0.25). The proportion of allantoin to total PD in urine ranged from 0.83 to 0.88, with the crossbreds having greater values compared with the 2 parents (P = 0.03). Daily glomerular filtration rate, calculated using endogenous creatinine as an internal marker, for the above animals was 3.85, 4.23, and 3.61 L/kg of BW(0.75), respectively, in fasting trial (P = 0.59). The alpine animals may develop special regulating mechanisms in the kidney in terms of glomerular filtration rate and PD excretion, which would help the animals in adapting to the harsh environment of the Qinghai-Tibetan plateau.