The biohydrogenation theory of milk fat depression (MFD) attributes decreases in milk fat in cows to the formation of specific fatty acids (FA) in the rumen.Trans-10,cis-12-CLA is the only biohydrogenation intermediate known to inhibit milk fat synthesis, but it is uncertain if increased ruminal synthesis is the sole explanation of MFD. Four lactating cows were used in a 4×4 Latin square with a 2×2 factorial arrangement of treatments and 35-d experimental periods to evaluate the effect of diets formulated to cause differences in ruminal lipid metabolism and milk fat synthesis on the flow of FA and dimethyl acetal at the omasum. Treatments comprised total mixed rations based on grass silage with a forage:concentrate ratio of 35:65 or 65:35 containing 0 or 50 g/kg sunflower oil (SO). Supplementing the high-concentrate diet with SO lowered milk fat synthesis from −20·2 to −31·9 % relative to other treatments. Decreases in milk fat were accompanied by alterations in ruminal biohydrogenation favouring thetrans-10 pathway and an increase in the formation of specific intermediates includingtrans-4 totrans-10-18 : 1,trans-8,trans-10-CLA,trans-9,cis-11-CLA andtrans-10,cis-15-18 : 2. Flow oftrans-10,cis-12-CLA at the omasum was greater on high- than low-concentrate diets but unaffected by SO. In conclusion, ruminaltrans-10,cis-12-CLA formation was not increased on a diet causing MFD suggesting that other biohydrogenation intermediates or additional mechanisms contribute to the regulation of fat synthesis in the bovine mammary gland.

New technologies related to the identification of bacterial communities in fresh forage and silage may give valuable detailed information on the best practices to produce animal feeds. The objective was to evaluate how management conditions during silage making manipulate the profile of bacterial communities and fermentation quality of grass silages. Silages were prepared from mixed timothy and meadow fescue grass using two compaction levels. As an additional treatment the grass was contaminated with soil and feces prior to tight compaction. Four additive treatments with different modes of action were applied: control without additive, formic acid-based additive, homofermentative lactic acid bacteria and salt-based additive. After 93 days the silos were opened, samples were taken and routinely analyzed. DNA extraction was carried out and PCR amplification of the bacterial 16S rRNA gene V4 region was performed using universal primers. The silage pH was higher for loose than tight compaction and higher for non-contaminated than for contaminated silages. Great shift was observed in bacterial profiles from fresh material towards silage. Lactobacillus genus was barely found on the relative abundance of fresh grass but became predominant in the final silage along with Sphingomonas genus. Use of additives improved fermentation quality and modified the bacterial profiles of grass ensiled under different management conditions.

The present study was conducted to investigate ruminal N metabolism in dairy cows using 15N labeled N sources [ammonia N (AN), soluble non-ammonia N (SNAN) from rapeseed meal, and insoluble non-ammonia N (NAN) from rapeseed meal]. To describe the observed pattern of 15N transactions in the rumen, dynamic compartmental models were developed. The experiment consisted of 3 experimental treatments allocated to 4 cows according to a changeover design. The results from 2 treatments (AN and rapeseed meal SNAN) are reported in this paper. Ammonia N and rapeseed SNAN, both labeled with 15N, were administered intraruminally. Rumen evacuations in combination with grab samples from the rumen contents were used to determine ruminal N pool sizes. The 15N-atom% excess was determined in N fractions of rumen digesta samples that were distributed between 0 and 82 h after dosing. For the AN treatment, a 2-compartment model was developed to describe the observed pattern in 15N-atom% excess pool sizes of AN and bacterial N and to estimate kinetic parameters of ruminal 15N transactions. For the SNAN treatment, an additional compartment of SNAN was included in the model. Model simulations were used to estimate N fluxes in the rumen. Both models described the observed pattern of 15N-atom% excess pool sizes accurately, based on small residuals between observed and predicted values. Immediate increases in 15N-atom% excess of bacterial N with AN treatment suggested that microbes absorbed AN from extracellular pools rapidly to maintain sufficient intracellular concentrations. Proportionally 0.69 of the AN dose was recovered as NAN flow from the rumen. A rapid disappearance of labeled SNAN from rumen fluid and appearance in bacterial N pool indicated that, proportionally, 0.56 of SNAN was immediately either adsorbed to bacterial cell surfaces or taken up to intracellular pools. Immediate uptake of labeled SNAN was greater than that of AN (proportionally 0.56 vs. 0.16 of the dose). Degradation rate of SNAN to AN was relatively slow (0.46/h), but only 0.08 of the SNAN dose was estimated to escape ruminal degradation because of rapid uptake by the bacteria. Overall, losses of the 15N dose as AN absorption and outflow from the rumen were higher (P < 0.01) for the AN than the SNAN treatment (0.31 and 0.11 of the dose, respectively). Consequently, recovery as NAN flow was greater for SNAN than for AN treatment (0.89 vs. 0.69 of the dose). Estimated rate of bacterial N recycling to AN was on average 0.006/h, which suggests that N losses due to intraruminal recycling are small in dairy cows fed at high intake levels. We conclude that SNAN isolated from rapeseed meal had better ruminal N utilization efficiency than AN, as indicated by smaller ruminal N losses as AN (0.11 vs. 0.31 of the dose) and greater bacterial N flow (0.81 vs. 0.69 of the dose). Furthermore, the current findings indicate that rapid adsorption of soluble proteins to bacterial cells plays an important role in ruminal N metabolism.

Four multiparous dairy cows were used in a 4 × 4 Latin square to examine how starch level and oil mixture impact dry matter (DM) intake and digestibility, milk yield and composition, rumen fermentation, ruminal methane (CH₄) emissions, and microbial diversity. Experimental treatments comprised high (HS) or low (LS) levels of starch containing 0 or 30 g of a mixture of sunflower and fish oils (2:1 w/w) per kg diet DM (LSO and HSO, respectively). Intake of DM did not differ between cows fed LS and HS diets while oil supplementation reduced DM intake. Dietary treatments did not affect milk and energy corrected milk yields. There was a tendency to have a lower milk fat concentration due to HSO compared with other treatments. Both high starch level and oil supplementation increased digestibility of gross energy. Cows receiving HS diets had higher levels of total rumen VFA while acetate was lower than LS without any differences in rumen pH, or ruminal CH₄ emissions. Although dietary oil supplementation had no impact on rumen fermentation, decreased CH₄ emissions (g/day and g/kg milk) were observed with a concomitant increase in Anoplodinium-Diplodinium sp. and Epidinium sp. but a decrease in Christensenellaceae, Ruminococcus sp., Methanobrevibacter ruminantium and Mbb. gottschalkii clades.

Grass was ensiled using an acid-based additive (AS) or homofermentative lactic acid bacteria (IS). In addition, barley grain was either dried (DB) or crimped and ensiled (EB). The feeds were fed as total mixed rations (TMR) in a 2 × 2 factorial arrangement to 16 Nordic Red dairy cows in four replicated Latin squares. The differences in the fermentation quality of the two grass silages were unexpectedly small. Dry matter intake was higher (p < 0.01) for cows fed AS than those fed IS but was not affected by barley preservation method. Ruminal molar proportion of butyrate tended to be higher in cows fed AS rather than IS (p < 0.10) in expense of acetate (p < 0.05). Barley preservation method did not affect rumen fermentation but modulated rumen bacterial community composition. Milk production was not affected by silage additive but tended (p < 0.10) to be higher (39.6 vs. 39.0 kg/d) for cows fed DB rather than EB. However, barley type did not affect energy corrected milk yield due to a tendency (p < 0.10) for higher milk fat content of cows fed EB rather than DB. Milk fat yield tended (p < 0.10) to be higher for AS-fed cows than IS-fed cows, and milk protein yield was higher for cows receiving DB rather than EB. The AS resulted in more aerobically stable TMR than IS and a minor advantage was found for DB compared to EB.

Accurate quantitative information on the fate of dietary protein in the rumen is central to modern metabolizable protein systems developed to improve the efficiency of nitrogen utilization in ruminants. An in vitro method was developed to estimate the rate of soluble rapeseed meal (Brassica rapa L.) protein (SRMP) degradation. Unlabeled and (15)N-labeled solvent-extracted rapeseed meal were incubated alone or as an equal mixture (125 mg of N/L) with buffered rumen contents and a mixture of carbohydrates formulated to provide a constant source of fermentable energy during the course of all incubations. Incubations were made over 0.33, 0.67, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 6.0, 8.0, and 10.0 h. Enrichment of (14)N and (15)N isotopes in total N of ammonia (AN), soluble nonammonia (SNAN), and insoluble (ISN) fractions liberated during incubations with test proteins was determined. A model with 4 pools that accounted for both intracellular and extracellular N transformations was used to estimate the rate of SRMP degradation. Parameter values used in the model were adjusted based on the size of A(14)N, A(15)N, SNA(14)N, SNA(15)N, IS(14)N, and IS(15)N pools, measured at different time points during incubations with buffered rumen fluid. The mean rate of N degradation for SRMP was estimated at 0.126 (SD 0.0499) h(-1). No substantive difference in the rate of protein degradation or microbial protein synthesis was observed during incubations of labeled and unlabeled substrates with rumen fluid. In conclusion, combined use of data from incubations of unlabeled and (15)N-labeled rapeseed protein with buffered rumen inoculum provided sufficient information to allow for estimation of parameter values in a complex dynamic model of soluble protein degradation. Results indicate the potential of the technique to evaluate the degradability of SNAN of other dietary protein sources and implicate ruminal escape of soluble rapeseed protein as an important source of amino acids in ruminants.