Carla R. Soliva

Efficacy of plant extracts rich in secondary constituents to modify rumen fermentation

Three plant extracts with different secondary constituents were simultaneously evaluated in an eight fermenter rumen simulation technique (Rusitec) for their effects on ruminal fermentation pattern. Specifically, a Yucca schidigera extract containing the saponin compound sarsaponin was supplied at three concentrations (1, 20 and 100 mg sarsaponin/kg dry matter (DM)) and was compared to a Castanea sativa wood extract containing hydrolysable tannins (0.5 and 2.5 g tannins/kg) and pure sulphonate-free lignin (2.5 g/kg). The supplements were added to a basal diet (grass silage, barley grain, grass hay) with a low crude protein (CP) content. These diets were compared to the same, unsuppplemented basal diet (control) and to a diet with additional soybean meal providing protein according to recommendations for dairy cow diets. The control diet and the soybean meal diet differed in rumen fluid ammonia concentration by 15% (13.6 mmol/l versus 16.0 mmol/l). Relative to the control diet, the high doses of the saponin-rich and the tannin-rich extract reduced the rumen fluid ammonia level by up to 21%. There was a tendency for elevated levels of apparent protein degradation during 48 h of fermentation, suggesting that the effect of the extracts on ammonia probably did not result from suppression of microbial protein degradation. Bacterial and protozoal counts in rumen fluid were not influenced by dietary treatments. Substantive effects of the saponin-rich and tannin-rich products on ruminal nitrogen metabolism were observed only at doses exceeding those recommended by the manufacturers.

Methane-suppressing effect of myristic acid in sheep as affected by dietary calcium and forage proportion.

The efficiency of myristic acid (14 : 0) as a feed additive to suppress CH4 emissions of ruminants was evaluated under different dietary conditions. Six sheep were subjected to a 6 x 6 Latin square arrangement. A supplement of non-esterified 14 : 0 (50 g/kg DM) was added to two basal diets differing in their forage:concentrate values (1 : 1.5 and 1 : 0.5), which were adjusted to dietary Ca contents of 4.2 and 9.0 g/kg DM, respectively. Comparisons were made with the unsupplemented basal diets (4.2 g Ca/kg DM). The 14 : 0 supplementation decreased (P<0.001) total tract CH4 release depending on basal diet type (interaction, P<0.001) and dietary Ca level (P<0.05, post hoc test). In the concentrate-based diet, 14 : 0 suppressed CH4 emission by 58 and 47 % with 4.2 and 9.0 g Ca/kg DM, respectively. The 14 : 0 effect was lower (22 %) in the forage-based diet and became insignificant with additional Ca. Myristic acid inhibited (P<0.05) rumen archaea without significantly altering proportions of individual methanogen orders. Ciliate protozoa concentration was decreased (P<0.05, post hoc test) by 14 : 0 only in combination with 9.0 g Ca/kg DM. Rumen fluid NH3 concentration and acetate:propionate were decreased (P<0.05) and water consumption was lower (P<0.01) with 14 : 0. The use of 14 : 0 had no clear effects on total tract organic matter and fibre digestion; this further illustrates that the suppressed methanogenesis resulted from direct effects against methanogens. The present study demonstrated that 14 : 0 is a potent CH4 inhibitor but, to be effective in CH4 mitigation feeding strategies, interactions with other diet ingredients have to be considered.

Effects of mixtures of lauric and myristic acid on rumen methanogens and methanogenesis in vitro

Aims: To identify the most effective mixture of non‐esterified lauric (C12) and myristic (C14) acid in suppressing ruminal methanogenesis, and to investigate their effects on the methanogenic population.

Rumen simulation technique study on the interactions of dietary lauric and myristic acid supplementation in suppressing ruminal methanogenesis.

The interactions of lauric (C12) and myristic acid (C14) in suppressing ruminal methanogenesis and methanogens were investigated with the rumen simulation technique (Rusitec) using bovine ruminal fluid. The fatty acids were added to basal substrates (grass hay:concentrate, 1:1.5) at a level of 48 g/kg DM, provided in C12:C14 ratios of 5:0, 4:1, 3:2, 2.5:2.5, 2:3, 1:4 and 0:5. Additionally, an unsupplemented control consisting of the basal substrates only was employed. Incubation periods lasted for 15 (n 4) and 25 (n 2) d. CH4 formation was depressed by any fatty acid mixture containing at least 40 % C12, and effects persisted over the complete incubation periods. The greatest depression (70 % relative to control) occurred with a C12:C14 ratio of 4:1, whereas the second most effective treatment in suppressing CH4 production (60 % relative to control) was found with a ratio of 3:2. Total methanogenic counts were decreased by those mixtures of C12 and C14 also successful in suppressing methanogenesis, the 4:1 treatment being most efficient (60 % decline). With this treatment in particular, the composition of the methanogenic population was altered in such a way that the proportion of Methanococcales increased and Methanobacteriales decreased. Initially, CH4 suppression was associated with a decreased fibre degradation, which, however, was reversed after 10 d of incubation. The present study demonstrated a clear synergistic effect of mixtures of C12 and C14 in suppressing methanogenesis, mediated probably by direct inhibitory effects of the fatty acids on the methanogens.

Palatability in sheep and in vitro nutritional value of dried and ensiled sainfoin (Onobrychis viciifolia) birdsfoot trefoil (Lotus corniculatus), and chicory (Cichorium intybus)

Abstract Three temperate forages, sainfoin, birdsfoot trefoil, and chicory, characterized by elevated contents of plant secondary compounds, were compared to a ryegrass-clover mixture (control) in dried (Experiment 1) and ensiled form (Experiment 2) in their palatability and nutritional value. Palatability was measured in adult wethers (n = 6) allowed to choose between the familiar control forage and one of the three test plants. Palatability index was calculated from differences in intake of control and test plants measured after given times. Generally at first contact, palatability of the unfamiliar plants was low. Lag time until palatability index approached or exceeded a value of 100 was 2 – 5 d, but could not be related to the content of condensed tannins. Sainfoin had a high palatability, the highest content of condensed tannins (77.4 ± 10.23 g/kg DM), a high content of duodenally utilisable crude protein (94.7 ± 16.87 g/100 g CP), and a high content of metabolizable energy (9.5 ± 0.38 MJ ME/kg DM), making this plant most promising for various purposes including anthelmintic action.

Ruminal methane inhibition potential of various pure compounds in comparison with garlic oil as determined with a rumen simulation technique (Rusitec).

Ruminants represent an important source of methane (CH(4)) emissions; therefore, CH(4) mitigation by diet supplementation is a major goal in the current ruminant research. The objective of the present study was to use a rumen simulation technique to evaluate the CH(4)-mitigating potential of pure compounds in comparison with that achieved with garlic oil, a known anti-methanogenic supplement. A basal diet (15 g DM/d) consisting of ryegrass hay, barley and soyabean meal (1:0·7:0·3) was incubated with the following additives: none (negative control); garlic oil (300 mg/l incubation liquid; positive control); allyl isothiocyanate (75 mg/l); lovastatin (150 mg/l); chenodeoxycholic acid (150 mg/l); 3-azido-propionic acid ethyl ester (APEE, 150 mg/l); levulinic acid (300 mg/l); 4-[(pyridin-2-ylmethyl)-amino]-benzoic acid (PABA, 300 mg/l). Fermentation profiles (SCFA, microbial counts and N turnover) and H(2) and CH(4) formation were determined. Garlic oil, allyl isothiocyanate, lovastatin and the synthetic compound APEE decreased the absolute daily CH(4) formation by 91, 59, 42 and 98 %, respectively. The corresponding declines in CH(4) emitted per mmol of SCFA were 87, 32, 40 and 99 %, respectively, compared with the negative control; the total SCFA concentration was unaffected. Garlic oil decreased protozoal numbers and increased bacterial counts, while chenodeoxycholic acid completely defaunated the incubation liquid. In vitro, neutral-detergent fibre disappearance was lower following chenodeoxycholic acid and PABA treatments (- 26 and - 18 %, respectively). In conclusion, garlic oil and APEE were extremely efficient at mitigating CH(4) without noticeably impairing microbial nutrient fermentation. Other promising substances were allyl isothiocyanate and lovastatin.

Methane output of rabbits (Oryctolagus cuniculus) and guinea pigs (Cavia porcellus) fed a hay-only diet: Implications for the scaling of methane production with body mass in non-ruminant mammalian herbivores

It is assumed that small herbivores produce negligible amounts of methane, but it is unclear whether this is a physiological peculiarity or simply a scaling effect. A respiratory chamber experiment was conducted with six rabbits (Oryctolagus cuniculus, 1.57±0.31 kg body mass) and six guinea pigs (Cavia porcellus, 0.79±0.07 kg) offered grass hay ad libitum. Daily dry matter (DM) intake and DM digestibility were 50±6 g kg⁻⁰·⁷⁵ d⁻¹ and 55±6% in rabbits and 59±11 g kg⁻⁰·⁷⁵ d⁻¹ and 61±3% in guinea pigs, respectively. Methane production was similar for both species (0.20±0.10 L d⁻¹ and 0.22±0.08L d⁻¹ and represented 0.69±0.32 and 1.03±0.29% of gross energy intake in rabbits and guinea pigs, respectively. In relation to body mass (BM) guinea pigs produced significantly more methane. The data on methane per unit of BM obtained in this study and from the literature on the methane output of elephant, wallabies and hyraxes all lay close to a regression line derived from roughage-fed horses, showing an increase in methane output with BM. The regression, including all data, was nearly identical to that based on the horse data only (methane production in horses [L d⁻¹]=0.18 BM [kg]⁰·⁹⁷(⁹⁵%CI ⁰·⁹²⁻¹·⁰²)) and indicates linear scaling. Because feed intake typically scales to BM⁰·⁷⁵, linear scaling of methane output translates into increasing energetic losses at increasing BM. Accordingly, the data collection indicates that an increasing proportion of ingested gross energy is lost because relative methane production increases with BM. Different from ruminants, such losses (1%-2% of gross energy) appear too small in non-ruminant herbivores to represent a physiologic constraint on body size. Nevertheless, this relationship may represent a physiological disadvantage with increasing herbivore body size.

Transfer of linoleic and linolenic acid from feed to milk in cows fed isoenergetic diets differing in proportion and origin of concentrates and roughages.

The transfer of ingested alpha-linolenic acid (ALA) and linoleic acid (LA) determines the nutritional quality of milk, but the factors determining this transfer are unclear. The present experiment investigated the influence of roughage to concentrate proportions and the effect of concentrate types on milk fat composition. Respectively, six lactating dairy cows were fed one of three isoenergetic (5.4+/-0.05 MJ net energy for lactation/kg dry matter; DM) and isonitrogenous (215+/-3.5 g crude protein/kg DM) diets, consisting of ryegrass hay only (33 g fatty acids/kg DM; ALA-rich, no concentrate), maize (straw, whole maize pellets and gluten; 36 g fatty acids/kg DM; LA-rich; 560 g concentrate/kg DM), or barley (straw and grain plus soybean meal; 19 g fatty acids/kg DM; LA-rich; 540 g concentrate/kg DM). The fatty acid composition of feeds and resulting milk fat were determined by gas chromatography. The ALA concentration in milk fat was highest (P<0.001) with the hay-diet, but the proportionate transfer of ALA from diet to milk was lower (P<0.001) than with the maize- or barley-diets. The LA concentration in milk fat was highest with the maize-diet (P<0.05, compared with hay) but relative transfer rate was lower (P=0.01). The transfer rates of ALA and LA were reciprocal to the intake of individual fatty acids which thus contributed more to milk fat composition than did roughage to concentrate proportions. The amount of trans-11 18:1 in milk fat was lowest with the barley-diet (P<0.001) and depended on the sum of ALA and LA consumed. The milk fat concentration of cis-9, trans-11 18:2 (rumenic acid) was more effectively promoted by increasing dietary LA (maize) than ALA (hay). Amounts of 18:0 secreted in milk were four (maize) to seven (hay) times higher than the amounts ingested. This was suggestive of a partial inhibition of biohydrogenation in the maize-diet, possibly caused by the high dietary LA level.

Experimental validation of the Intergovernmental Panel on Climate Change default values for ruminant-derived methane and its carbon-isotope signature.

Two aspects regarding the ruminants contribution to global methane (CH4) emissions were investigated: (i) testing the accuracy of the Intergovernmental Panel on Climate Change default values for dairy cows fed different diet types and differing slurry storage temperatures; and (ii) providing carbon-isotope (C-isotope) signature data to contribute information on the characteristics of ruminant-derived CH4 as global source. The experimental diets, fed to 18 dairy cows, were separated into forage-only (hay, C3 plant) and forage-concentrate diets (barley, C3 plant; maize, C4 plant). Accumulated slurry was stored at either 14 or 27C. The hay diet had the highest CH4 conversion rate (Ym 7.9%). Negligible amounts of CH4 were emitted from slurries stored at low temperature. No diet effect was found at 27C (~33 L/kg volatile solids). The isotope ratios of enteric CH4 averaged 67.7‰ (C3 plants) and 57.4‰ (C4; maize). High temperature slurry storage resulted in different enrichment factors eCO2-CH4 for maize (33.2‰) and hay (35.9‰). Compared with the Intergovernmental Panel on Climate Change default values for Ym and slurry CH4 emission the results gained in the present experiment were higher and lower, respectively. Slurry-derived CH4 was less depleted in 13 C than

Fatty acid profile and oxidative stability of the perirenal fat of bulls fattened on grass silage and maize silage supplemented with tannins, garlic, maca and lupines

Carcass fat composition of cattle fed a forage-based diet might be inferior with maize silage compared to grass-silage based systems. This was quantified using complete diets with concentrate. To test whether supplements may influence carcass fat properties as well, the maize-silage diet was additionally supplemented either with Acacia mearnsii tannins, garlic, maca or lupines, feeds rich in secondary metabolites. The perirenal fat of 6×6 bulls fed these six diets was analysed for fatty acid profile and shelf life. The n-6/ n-3 ratio was always higher than 11 with the maize silage treatments and 2 with grass silage. The supplements did not affect the occurrence of biohydrogenation intermediates, including rumenic acid. Shelf life, being twice as long with maize compared to grass silage, was either unaffected or tended to be impaired, especially with supplementary garlic. Overall, supplementation was not efficient in improving carcass fat properties of maize-silage fed bulls.