Methods of Emulsifying Linoleic Acid in Biohydrogenation Studies In Vitro May Bias the Resulting Fatty Acid Profiles

The effects of three emulsifying methods on ruminal fatty acid biohydrogenation (BH) in vitro were compared. Using a static in-vitro gas test system, four replicates of each treatment were incubated in buffered ruminal fluid. Hemicellulose (300mg dry matter) was supplemented either with or without linoleic acid (9c12c-18:2, 5% in diet dry matter) and incubated for 4 and 24h. Three methods of emulsifying 9c12c-18:2 were tested: (1) ethanol, (2) Tween® 80, and (3) sonication. The products were then compared to non-emulsified 9c12c-18:2. Out of the three emulsifying methods tested, ethanol and sonication resulted in stable 9c12c-18:2 emulsions, indicating good 9c12c-18:2 distribution, while the Tween® 80 emulsion was less stable. BH was strongly inhibited by treating 9c12c-18:2 with ethanol and sonication at different steps of the BH-pathway, resulting in changed concentrations of certain BH intermediates. The fatty acid profile generated from the major BH-pathways of 9c12c-18:2 with Tween® 80 was comparable to that without emulsification after 24h of incubation. We conclude that it is not recommended to emulsify lipids before incubating them in vitro when investigating fatty acid BH. If emulsification of 9c12c-18:2 is necessary, Tween® 80 seems to be the method that interferes least with B

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 × 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:pro-pionate 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 considere

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 methanogen

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 (CH4) emissions; therefore, CH4 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 CH4-mitigating potential of pure compounds in comparison with that achieved with garlic oil, a known anti-methanogenic supplement. A basal diet (15g 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 (300mg/l incubation liquid; positive control); allyl isothiocyanate (75mg/l); lovastatin (150mg/l); chenodeoxycholic acid (150mg/l); 3-azido-propionic acid ethyl ester (APEE, 150mg/l); levulinic acid (300mg/l); 4-[(pyridin-2-ylmethyl)-amino]-benzoic acid (PABA, 300mg/l). Fermentation profiles (SCFA, microbial counts and N turnover) and H2 and CH4 formation were determined. Garlic oil, allyl isothiocyanate, lovastatin and the synthetic compound APEE decreased the absolute daily CH4 formation by 91, 59, 42 and 98%, respectively. The corresponding declines in CH4 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 CH4 without noticeably impairing microbial nutrient fermentation. Other promising substances were allyl isothiocyanate and lovastati

Stable Carbon Isotope Composition of c 9, t 11-Conjugated Linoleic Acid in Cow's Milk as Related to Dietary Fatty Acids

This study explores the potential use of stable carbon isotope ratios (δ13C) of single fatty acids (FA) as tracers for the transformation of FA from diet to milk, with focus on the metabolic origin of c9,t11-18:2. For this purpose, dairy cows were fed diets based exclusively on C3 and C4 plants. The FA in milk and feed were fractionated by silver-ion thin-layer chromatography and analyzed for their δ13C values. Mean δ13C values of FA from C3 milk were lower compared to those from C4 milk (−30.1‰ vs. −24.9‰, respectively). In both groups the most negative δ13C values of all FA analyzed were measured for c9,t11-18:2 (C3 milk=−37.0±2.7‰; C4 milk −31.4±1.4‰). Compared to the dietary precursors 18:2n-6 and 18:3n-3, no significant 13C-depletion was measured in t11-18:1. This suggests that the δ13C-change in c9,t11-18:2 did not originate from the microbial biohydrogenation in the rumen, but most probably from endogenous desaturation of t11-18:1. It appears that the natural δ13C differences in some dietary FA are at least partly preserved in milk FA. Therefore, carbon isotope analyses of individual FA could be useful for studying metabolic transformation processes in ruminant

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 α-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 leve

Efficiency of monolaurin in mitigating ruminal methanogenesis and modifying C-isotope fractionation when incubating diets composed of either C3 or C4 plants in a rumen simulation technique (Rusitec) system

Mitigation of methanogenesis in ruminants has been an important goal for several decades. Free lauric acid, known to suppress ruminal methanogenesis, has a low palatability; therefore, in the present study the aim was to evaluate the mitigation efficacy of its esterified form (monolaurin). Further, 13C-isotope abundance (δ13C) and 13C-12C fractionation during methanogenesis and fermentation were determined to evaluate possible microbial C-isotope preferences. Using the rumen simulation technique, four basal diets, characterised either by the C3 plants grass (hay) and wheat (straw and grain), or the C4 plant (13C excess compared with C3 plants) maize (straw and grain), and a mixture of the latter two, were incubated with and without monolaurin (50g/kg dietary DM). Added to hay, monolaurin did not significantly affect methanogenesis. When added to the other diets (P<0·05 for the wheat-based diet) methane formation was lowered. Monolaurin decreased fibre disappearance (least effect with the hay diet), acetate:propionate ratio, and protozoal counts. Feed residues and SCFA showed the same δ13C as the diets. Methane was depleted in 13C while CO2 was enriched in 13C compared with the diets. Monolaurin addition resulted in 13C depletion of CO2 and enrichment in CH4 (the latter only in the hay diet). In conclusion, monolaurin proved to effectively decrease methanogenesis in the straw-grain diets although this effect might partly be explained by the concomitantly reduced fibre disappearance. The influence on 13C-isotope abundance and fractionation supports the hypothesis that ruminal microbes seem to differentiate to some extent between C-isotopes during methanogenesis and fermentatio

Akzeptanz von Futterpflanzen mit Vorkommen von kondensierten Tanninen bei Schafen und ihre Gehalte an nutzbarem Rohprotein

Plants with moderate contents of condensed tannins are assumed to have anthelmintic Plants with moderate contents of condensed tannins are assumed to have anthelmintic properties and to improve protein supply of ruminants by reducing its ruminal degradability. In the present study, birdsfoot trefoil (Lotus corniculatus), chicory (Cychorium intybus) and sainfoin (Onobrychis viciifolia), suitable to be grown in Central Europe, were tested for their suitability as feed for ruminants. Sainfoin proved to be the most promising forage plant since it was very well accepted by the sheep and presented the highest content of utilizable crude protein as determined in vitro. Only few condensed tannins were detected in chicory. Further in vivo studies are required to confirm a better metabolic protein supply of ruminants when fed sainfoin

Methane Output of Tortoises: Its Contribution to Energy Loss Related to Herbivore Body Mass

An increase in body mass (M) is traditionally considered advantageous for herbivores in terms of digestive efficiency. However, recently increasing methane losses with increasing M were described in mammals. To test this pattern in non-mammal herbivores, we conducted feeding trails with 24 tortoises of various species (M range 0.52–180 kg) fed a diet of grass hay ad libitum and salad. Mean daily dry matter and gross energy intake measured over 30 consecutive days scaled to M0.75 (95%CI 0.64–0.87) and M0.77 (95%CI 0.66–0.88), respectively. Methane production was measured over two consecutive days in respiration chambers and scaled to M1.03 (95%CI 0.84–1.22). When expressed as energy loss per gross energy intake, methane losses scaled to 0.70 (95%CI 0.47–1.05) M0.29 (95%CI 0.14–0.45). This scaling overlaps in its confidence intervals to that calculated for nonruminant mammals 0.79 (95%CI 0.63–0.99) M0.15 (95%CI 0.09–0.20), but is lower than that for ruminants. The similarity between nonruminant mammals and tortoises suggest a common evolution of the gut fauna in ectotherms and endotherms, and that the increase in energetic losses due to methane production with increasing body mass is a general allometric principle in herbivores. These findings add evidence to the view that large body size itself does not necessarily convey a digestive advantage