Biohydrogenation Of Fatty Acids Research Articles

Changes in the fatty acid composition of steer subcutaneous fat, including biohydrogenation products, are minimal when finished on combinations of corn and barley grains and silages

Corn production in Western Canada has increased with development of short-season varieties partially offsetting the use of barley as a silage and grain source. The current study evaluated effects of silage and cereal-grain source on subcutaneous fat (SCF) composition of finishing cattle. Steers (465 ± 28.0 kg) were assigned to 24 pens (12 steers per pen) in a 2 × 3 factorial design for an 89 d finishing study. Diets contained corn silage or barley silage at 8% [dry matter (DM)] combined either dry-rolled barley grain (BG; 86% of DM), corn grain (CG; 85% of DM), or an equal blend of BG and CG (85% of DM). Bone-in ribeyes were collected from four steers per pen, and subcutaneous fatty acids were analyzed by gas chromatography. Feeding dry-rolled CG increased dietary fat and 18:2n-6 concentrations, resulting in small increases in 18:2n-6, total n-6 fatty acids, and n-6/n-3 ratios in SCF (P < 0.05). There were no changes in proportions of major trans-18:1 isomers (t10-18:1 or t11-18:1), the main natural isomer of conjugated linoleic acid (cis9,t11–18:2), or n-3 fatty acids. Substitution of dry-rolled CG for BG in finisher diets can, therefore, be done without substantially altering the fatty acid composition of beef fat, including polyunsaturated fatty acid biohydrogenation products.

Ruminal parameters and fatty acid composition of omasal digesta and milk in cows fed sugarcane-based diets supplemented with sunflower oil

This study evaluates the intake and digestion of nutrients, parameters of rumen fermentation and degradation, omasal digesta and milk fatty acid composition, productive performance, and the concentration of serum metabolites in cows fed 600 g kg-1 sugarcane-based diets containing 0 (control), 15, 30, and 45 g kg-1 sunflower oil (SO) on a dry matter (DM) basis. Four rumen-cannulated Holstein x Gyr cows yielding 15±5 kg day-1 with 110±10 days in milk were allocated in a 4 x 4 Latin square design. Data were analyzed using mixed models, and significant differences were declared at P < 0.05. There was no effect of SO on the intake and apparent digestibility of DM, crude protein, neutral detergent fiber (NDF) and nonfibrous carbohydrates, but there was a linear increase in the intake and digestibility of ether extract. Dietary SO levels did not alter the ruminal degradability parameters for DM and NDF, rumen pH and contents of ammonia N, acetate, propionate and volatile fatty acids. Milk fat content and yield were linearly decreased, whereas a linear increase in milk protein content was observed in response to increasing levels of SO, but with no effect on milk yield. Linear reductions in palmitic and ?-linolenic acid contents, a linear increase in trans-10 C18:1 and elaidic acids, and a quadratic effect on vaccenic and rumenic acids were observed in omasal digesta of cows fed increasing levels of SO. Overall, up to 45 g kg-1 SO can be included on DM of chopped sugarcane-based diets without reducing consumption, apparent digestibility and rumen degradability of DM and fiber. Supplementing chopped sugarcane-based diets with 30 to 45 g kg-1 SO (DM basis) promotes milk fat depression due to the inhibition of mammary lipogenesis by specific rumen-derived fatty acid intermediates of the biohydrogenation of unsaturated C18 fatty acids. The inclusion of 15 to 45 g kg-1 SO in chopped sugarcane-based diets improves the nutritional quality of milk fat, with increases in the levels of oleic, vaccenic and rumenic acids, beneficial to human health, and a reduction in the levels of the hypercholesterolemic lauric, myristic and palmitic acids.

Effect of toasting and decortication of oat on rumen biohydrogenation and intestinal digestibility of fatty acids in dairy cows

This experiment quantified the effect of decorticated and toasted oat (Avena sativa L.) on fatty acid (FA) supply, ruminal biohydrogenation (BH) of FA, and intestinal digestibility of FA in 4 ruminal and intestinal cannulated Danish Holstein cows. Experimental diets containing untreated oat, decorticated oat, toasted oat, and decorticated and toasted oat were fed ad libitum to the cows in a 2 × 2 factorial arrangement in a Latin square design throughout 4 periods. Unless otherwise mentioned, the results of this study indicate the main effect of decortication and toasting. Decortication increased the intake of FA by 40.3 g/d and increased feed-ileum digested FA, whereas toasting decreased the intake of FA by 69.3 g/d. Toasting increased both feed-ileum and total-tract digestibility of FA by 59.8 and 67.4 g/kg of FA intake, respectively. The proportion of C18:2n-6 in FA intake increased, and the C18:3n-3 proportion in FA intake decreased due to decortication. Toasting resulted in a dramatic reduction of the C18:2n-6 proportion in FA intake, and it increased the proportions of C18:0 and C18:3n-3 in FA intake. Toasting reduced ruminal BH of C18:1n-9 and C18:2n-6 by 134 and 11.7 g/kg of FA intake, respectively, and toasting increased the proportion of unsaturated FA to saturated FA in the duodenal FA flow. Decortication decreased the ruminal BH of C18:3n-3 by 38.0 g/kg of FA intake. Decortication increased small intestinal digestibility of C12:0, C15:0, C20:0, and C22:0. Toasting increased the small intestinal digestibility of C15:0, C18:0, trans-C18:1, C20:0, and C24:0. Toasting reduced the small intestinal digestibility of C18:1n-9, C18:2n-6, and C20:1n-9. This study showed that decortication successfully increased the intake of FA and flow of FA at the duodenum and feed-ileum digested FA. However, toasting oat at 121°C caused a remarkable decline in FA concentration in oat, and thereby FA intake; therefore, toasting cannot be recommended.

In vitro ruminal biohydrogenation of C18 fatty acids in mixtures of Indigofera zollingeriana and Brachiaria decumbens

This research was aimed at studying the in vitro ruminal biohydrogenation (BH) of C18 fatty acids (FA) in mixtures of Indigofera zollingeriana and Brachiaria decumbens. Four combinations of experimental rations of I. zollingeriana : B. decumbens were tested i.e., IZ 1 (45%:45%), IZ 2 (60%:30%), IZ 3 (75%:15%), and IZ 4 (90%:0%). The remaining 10% in in each ration was rice bran. The experimental design was based on a completely randomized design with five replicates. Results revealed that there was a statistically significant difference (P<0.01) in the composition of C18 unsaturated FA (UFA) and saturated FA (SFA) for each in vitro incubation period of 1, 2, 4, 8 and 24 h. The highest accumulation of C18 UFA at 24 h was observed in the incubation of IZ 4 (19.87%). The BH of C18:3, C18:2, and C18:1 showed no differences (P>0.05). Composition of C18:0 after incubation showed a significant difference (P<0.01) with the lowest composition was observed in IZ 2 (22%). In conclusion, combination of I. zollingeriana and B. decumbens at different ratio has minor inhibition effect on BH of C18 UFA.

Effects of dietary flaxseed and vitamin E on fermentation, nutrient disappearance, fatty acid biohydrogenation, and microbial protein synthesis using a simulated rumen (Rusitec)

Two simulated rumens (Rusitecs) were used to assess the effects of flaxseed (FS) and (or) vitamin E (VE) on rumen fermentation, fatty acid (FA) biohydrogenation, and microbial protein synthesis. Ground FS replaced 0% or 15% of barley grain, along with VE at 0 or 1000 IU d−1 in a 2 × 2 factorial experiment. Flaxseed lowered neutral detergent fiber (P = 0.001) and acid detergent fiber (P = 0.01) and increased (P = 0.001) nitrogen (N) disappearance. Flaxseed also increased (P = 0.01) total volatile FA and decreased (P = 0.001) acetate production. When both FS and VE were included, the acetate:propionate ratio decreased (P = 0.04). Biohydrogenation of FA was not influenced by VE, but total FA and C18:0 in effluent were increased (P = 0.001) and C16:0 decreased (P = 0.001) by FS. With VE, total microbial N (MN) was increased (P = 0.001). In the concentrate, production of MN in feed-particle-bound bacteria was increased (P = 0.001) by VE. Vitamin E did not alter FA biohydrogenation but did promote MN production. The stable and relatively high pH in the Rusitec may have prevented the typical shift from C18:1 trans-11 to C18:1 trans-10 with concentrate diets. Future studies simulating subclinical acidosis in the Rusitec may illuminate ruminal mode(s) of action of VE on FA biohydrogenation.

In vitro gas production and rumen fermentation profile of fresh and ensiled genetically modified high–metabolizable energy ryegrass

We previously generated a high-metabolizable energy (HME) perennial ryegrass (Lolium perenne) by genetically modifying the plant to increase the leaf lipid content. Although substantial progress has been made toward characterizing physiological changes of HME ryegrass, very limited information exists for feeding value and its suitability for adoption into the pastoral system. In this study, independent HME ryegrass lines with a range of elevated leaf lipid concentrations were analyzed for changes in fatty acids and possible associated changes in the broader nutritional profile, including the gross energy, which was found to increase by 6.8%. Because ryegrass is often ensiled and fermentation in the rumen leads to biohydrogenation of fatty acids as well as enteric methane production, we sought to investigate these effects on HME ryegrass. This was achieved by performing mini-scale silos and using an automated gas measurement system to incubate the material in rumen fluid in vitro for 24 h. Our study included treatments comprising 3 independent HME ryegrass genotypes and wild-type control materials prepared fresh and as silage, employing in total 5 incubation studies, using rumen fluids collected from 4 nonlactating Jersey × Holstein cows. At intervals during the incubation, the production of gases, volatile fatty acids, and the degree of biohydrogenation were measured. Statistical data analysis indicated that differences in the nutritional compositions of the ensiled materials largely reflected those of their fresh counterparts. Incubation of both fresh and ensiled HME ryegrass in rumen fluid resulted in: (1) a greater percentage of valuable unsaturated fatty acids compared with the control; (2) a significant reduction of butyrate; and (3) a 10 to 15% decrease in the methane proportion of the total gas production. We conclude that ensiling could be a convenient option for preserving HME as a locally produced high-value supplementary feed; however, large-scale application needs to be investigated. In this paper we discuss the potential use of HME ryegrass to enhancing forage feeding value and the potential environmental benefits to the pastoral agriculture industry.

Potential Health Benefits of Conjugated Linoleic Acid: An Important Functional Dairy Ingredient

Conjugated linoleic acid (CLA) refers to a class of positional and geometrical isomers of linoleic acid (cis-9, cis-12 octadecadienoic acid) having conjugate double bond system. CLA are synthesized in rumen of the ruminants by biohydrogenation of dietary fatty acids; and thus, can be obtained from dairy products as well as from the meat of sheep, lamb and other ruminants. Among the several isomers, c9, t11-CLA isomer is the most biologically active form and accounts approximately 80% of total isomers. A number of clinical and epidemiological studies have demonstrated the role of CLA as anti-atherogenic, anti-inflammatory, anti-oxidative, anti-carcinogenic, etc. Several researchers have suggested the positive association of CLA in weight management, hypercholesterolemia, immunomodulatory functions, and improved bone metabolism.

In vitro rumen biohydrogenation of unsaturated fatty acids in tropical grass-legume rations.

Aim:The aim of this study was to evaluate the effects of various combinations of tropical grass-legume species in rations on the biohydrogenation (BH) activity of unsaturated fatty acids (FAs), C18:0 composition, and fermentation profile in an in vitro rumen system.Materials and Methods:Samples of the following five fodder plants were used: One species of grass (Pennisetum purpureum) and four species of tree legumes (Leucaena leucocephala, Gliricidia sepium, Calliandra calothyrsus, and Indigofera zollingeriana). The following eight experimental diets were evaluated: 50% P. purpureum + 50% L. leucocephala (LL I); 50% P. purpureum + 50% G. sepium (GS I); 50% P. purpureum + 50% C. calothyrsus (CC I); 50% P. purpureum + 50% I. zollingeriana (IZ I); 75% P. purpureum + 25% L. leucocephala (LL II); 75% P. purpureum + 25% G. sepium (GS II); 75% P. purpureum + 25% C. calothyrsus (CC II); and 75% P. purpureum + 25% I. zollingeriana (IZ II). Each ration was replicated 3 times. In vitro rumen incubation was performed for 48 h, according to the Tilley and Terry method. Determination of the FA profiles of the forage materials and rumen fluid samples was performed using gas chromatography.Results:The percentage of polyunsaturated FA (PUFA) in the forage materials ranged from 34.18% (P. purpureum) to 74.51% (C. calothyrsus). The percentage of monounsaturated FA (MUFA) ranged from 5.06% (P. purpureum) to 8.71% (L. leucocephala). The percentage of saturated FA (SFA) was the lowest at 19.12% (C. calothyrsus) and highest at 60.76% (P. purpureum). In vitro BH of C18:3 n-3, C18:2 n-6, C18:1 n-9, and C18 PUFA in the experimental diets ranged from 72% to 100%. The BH of C18:1 n-9 in GS I (80%) and IZ I (72%) was significantly different (p<0.05). The percentage of C18:0 was 10-50% and significantly different (p<0.05) among treatments, with the highest (of 50%) in GS II. No significant differences (p>0.05) were observed in the fermentation parameters (pH, total volatile FAs, in vitro dry matter digestibility, and in vitro organic matter digestibility) among the treatments, except in NH3 concentration (p<0.05).Conclusion:The various combinations of tropical legumes do not have significant inhibitory effects on the BH of C18:2 n-6, C18:3 n-3, and C18 PUFA after in vitro incubation for 48h. Furthermore, an increase in the tropical legume ratio in the ration tends to suppress C18:0 formation after the fermentation simulation process. IZ I has the potential to reduce C18:1 n-9 (MUFA) disappearance and yield an ideal rumen fermentation profile.

Effects of replacing starch with three sugars in a concentrate and forage diet on in vitro rumen fermentation, fatty acid composition and related bacteria

Context Replacing starch with sugar could maintain dietary energy density with reduced risks of rumen acidosis and milk fat depression, but the underlying mechanism is not well understood, and the effects of sugar feeding might vary among sugars. Aims Objectives of the present study were to evaluate the effects of replacing corn starch in a diet containing 40:60 forage-to-concentrate ratio (control) with 3%, 6% and 9% of sucrose, fructose and lactose on in vitro rumen fermentation, fatty acid (FA) composition and populations of bacteria involved in the production of trans-11 and trans-10 FA. Methods A 3 × 3 + 1 (control) factorial experimental design was used, and the pH, concentrations of volatile fatty acids (VFA) and ammonia-N, profiles of FA and the relative abundance of four trans-11-producing bacteria and two trans-10 FA-producing bacteria were measured after a 6-h incubation. Key results Replacing dietary corn starch with sucrose, fructose and lactose neither altered the concentration of total VFA after 6-h fermentation, nor decreased the pH, except for substitution with 9% sucrose. Increased butyrate proportions and decreased branched-chain VFA proportions were the common effects in sugar treatments, but the proportions of acetate and propionate varied among sugars. Lactose inclusion in the diet led to a higher pH, greater acetate and butyrate concentrations, and lower propionate concentrations than did sucrose inclusion. Sugar substitution decreased the concentrations of C18:1 trans-4 and most C18:2 isomers, but did not influence the major isomers related to trans-11 and trans-10 biohydrogenation pathways. Abundance of the four measured trans-11 FA-producing bacteria was increased by sugars, with sucrose showing a greater influence than did fructose and lactose. As to trans-10 FA-producing bacteria, only Megasphaera elsdenii populations were decreased by 3% and 6% fructose inclusion compared with the control. Dose-effect varied among sugars and the parameters measured, with sucrose having the most obvious dose effect among the three sugars; however, fructose affected mainly fermentation parameters, while lactose affected mainly C18 FA profiles. Conclusions Replacing corn starch in a high-concentrate diet with up to 9% of sucrose, fructose and lactose differentially affected rumen fermentation and rumen FA metabolism, by influencing the abundance of rumen bacteria involved in rumen FA biohydrogenation. Implications Lactose may be more efficient in increasing milk fat than are sucrose and fructose, and dose effect should be considered in the utilisation of sucrose.

390 Effects of dietary-induced pH changes on feed digestion, nitrogen metabolism, fermentation, and biohydrogenation of polyunsaturated fatty acids in dual-flow continuous culture fermenters

Abstract Ruminal pH decreases as greater concentrations of grain are consumed, and as a result, reduces biohydrogenation of polyunsaturated fatty-acids. Our objective was to investigate effects of diet on pH and biohydrogenation activity in vitro. Eight dual-flow, continuous culture fermenters were blocked by side of the room and randomly assigned, within block, to 1 of 4 treatments. Treatments consisted of 100:0 roughage-to-concentrate (Grass), 50:50 roughage-to-concentrate plus flax oil supplement (Mix+F), 10:90 roughage-to-concentrate plus corn oil supplement (Con+C), or 10:90 roughage-to-concentrate plus flax oil supplement (Con+F). Fermenter pH, area under the curve, and maximum pH were lowest for Con+C and Con+F (P &amp;lt; 0.05). Apparent and true DM, OM, and NDF digestibility were greater with grain inclusion (P &amp;lt; 0.05) while ADF digestibility was not affected by treatment (P &amp;gt; 0.10). Protein degradation was lowered as grain inclusion increased (P &amp;lt; 0.05). Total VFA concentrations were lowest for Grass, highest for Con+C and Con+F, and intermediate for Mix+F (P &amp;lt; 0.05). Acetate-to-propionate was lowest for Con treatments (P &amp;lt; 0.05). Biohydrogenation of 18:2n-6 and 18:3n-3 fatty-acids were reduced (P &amp;lt; 0.01) with inclusion of concentrate regardless of roughage-to-concentrate ratio. Concentrations of stearic acid recovered from effluent were greatest for Grass, intermediate for Mix+F, and lowest for both Con+C and Con+F (P &amp;lt; 0.05). Linoleic acid proportions in effluent were greatest for Con+C and Con+F and intermediate for Mix+F (P &amp;lt; 0.05), and inclusion of concentrate, regardless of amount, increased the amount linoleic acid recovered from fermenter overflow (P &amp;lt; 0.01). Between Con+C and Con+F concentrations of alpha-linolenic acid (ALA) was greater when flax oil was supplemented (P &amp;lt; 0.01). Proportions of ALA in effluent was similar (P &amp;gt; 0.10) between Grass and Con+C; however, between Mix+F and Con+F ALA concentrations tended (P = 0.06) to be greater for Con+F. Omega6-to-omega3 ratios were similar across Grass, Mix+F, and Con+F (P &amp;gt; 0.10) and greatest for Con+C (P &amp;lt; 0.05).

Corn oil supplementation enhances hydrogen use for biohydrogenation, inhibits methanogenesis, and alters fermentation pathways and the microbial community in the rumen of goats

Enteric methane (CH4) emissions are not only an important source of greenhouse gases but also a loss of dietary energy in livestock. Corn oil (CO) is rich in unsaturated fatty acid with >50% PUFA, which may enhance ruminal biohydrogenation of unsaturated fatty acids, leading to changes in ruminal H2 metabolism and methanogenesis. The objective of this study was to investigate the effect of CO supplementation of a diet on CH4 emissions, nutrient digestibility, ruminal dissolved gases, fermentation, and microbiota in goats. Six female goats were used in a crossover design with two dietary treatments, which included control and CO supplementation (30 g/kg DM basis). CO supplementation did not alter total-tract organic matter digestibility or populations of predominant ruminal fibrolytic microorganisms (protozoa, fungi, Ruminococcus albus, Ruminococcus flavefaciens, and Fibrobacter succinogenes), but reduced enteric CH4 emissions (g/kg DMI, -15.1%, P = 0.003). CO supplementation decreased ruminal dissolved hydrogen (dH2, P < 0.001) and dissolved CH4 (P < 0.001) concentrations, proportions of total unsaturated fatty acids (P < 0.001) and propionate (P = 0.015), and increased proportions of total SFAs (P < 0.001) and acetate (P < 0.001), and acetate to propionate ratio (P = 0.038) in rumen fluid. CO supplementation decreased relative abundance of family Bacteroidales_BS11_gut_group (P = 0.032), increased relative abundance of family Rikenellaceae (P = 0.021) and Lachnospiraceae (P = 0.025), and tended to increase relative abundance of genus Butyrivibrio_2 (P = 0.06). Relative abundance (P = 0.09) and 16S rRNA gene copies (P = 0.043) of order Methanomicrobiales, and relative abundance of genus Methanomicrobium (P = 0.09) also decreased with CO supplementation, but relative abundance (P = 0.012) and 16S rRNA gene copies (P = 0.08) of genus Methanobrevibacter increased. In summary, CO supplementation increased rumen biohydrogenatation by facilitating growth of biohydrogenating bacteria of family Lachnospiraceae and genus Butyrivibrio_2 and may have enhanced reductive acetogenesis by facilitating growth of family Lachnospiraceae. In conclusion, dietary supplementation of CO led to a shift of fermentation pathways that enhanced acetate production and decreased rumen dH2 concentration and CH4 emissions.

FATTY ACIDS PROFILE OF FRESH CHEESE PRODUCED IN AN INTENSIVE SILVOPASTORAL SYSTEM IN THE TROPICS

&lt;p&gt;Intensive silvopastoral systems are being increasingly disseminated in the tropical areas of Mexico, Central and South America, so it is important to know the quality of the animal products generated in these systems. Fatty acid profile of fresh soft cheese (Domiati type) produced from milk of cows grazing &lt;em&gt;Leucaena leucocephala&lt;/em&gt; associated with &lt;em&gt;Cynodon nlemfuensis&lt;/em&gt; in an intensive silvopastoral system (ISS) was evaluated and compared with a cheese produced from cows grazing a traditional monoculture system (MS) of &lt;em&gt;C. nlemfuensis&lt;/em&gt;. The legume &lt;em&gt;L. leucocephala&lt;/em&gt; contains phenolic compounds that could affect the fatty acid biohydrogenation in the rumen and therefore the fatty acid profile of resultant milk and cheese. Milk samples from cows in both diets (systems) were processed monthly for cheese making and cheese samples were analyzed for fatty acid profile. The experimental period lasted for 10 months covering the rainy and dry season of a tropical area in Mexico. The results showed that the cheese produced in ISS had significantly lower content (P&amp;lt;0.05) of the saturated fatty acids lauric, myristic and palmitic and significantly higher content (P&amp;lt;0.05) of stearic, linoleic and γ-linolenic acids. Additionally, higher content (P&amp;lt;0.05) of the polyunsaturated fatty acids and omega-6 fatty acids were found in the cheese from ISS. The results suggest that fresh soft cheese produced from silvopastoral systems based on &lt;em&gt;L. leucocephala&lt;/em&gt; has a better fatty acid profile than that produced from the traditional MS with grass only.&lt;/p&gt;

Effect of Feeding Cold-Pressed Sunflower Cake on Ruminal Fermentation, Lipid Metabolism and Bacterial Community in Dairy Cows.

Simple SummaryThe use of cold-pressed sunflower cake, a by-product of small-scale biodiesel manufacturing, as a substitute for prilled palm fat in dairy cows’ diet, can reduce the extent of unsaturated fatty acid biohydrogenation This favors an accumulation of vaccenic acid in the rumen concomitant with a greater daily duodenal microbial N flow and all without impairing ruminal fermentation, microbial diversity or abundance of dominant populations. In the present study, only changes in relative abundances of less-representative genera were induced.Cold-pressed sunflower cake (CPSC), by-product of oil-manufacturing, has high crude fat and linoleic acid concentrations, being a promising supplement to modulate rumen fatty acid (FA) profile. This trial studied CPSC effects on ruminal fermentation, biohydrogenation and the bacterial community in dairy cows. Ten cows were used in a crossover design with two experimental diets and fed during two 63-day periods. The cows were group fed forage ad libitum and the concentrate individually. The concentrates, control and CPSC, were isoenergetic, isoproteic and isofat. The ruminal samples collected at the end of each experimental period were analyzed for short-chain fatty acid, FA and DNA sequencing. CPSC decreased butyrate molar proportion (4%, p = 0.005). CPSC decreased C16:0 (28%, p < 0.001) and increased C18:0 (14%, p < 0.001) and total monounsaturated FA, especially C18:1 trans-11 (13%, p = 0.023). The total purine derivative excretion tended to be greater (5%, p = 0.05) with CPSC, resulting in a 6% greater daily microbial N flow. CPSC did not affect the diversity indices but increased the relative abundances of Treponema and Coprococcus, and decreased Enterococcus, Ruminococcus and Succinivibrio. In conclusion, the changes in ruminal fermentation and the FA profile were not associated with changes in microbial diversity or abundance of dominant populations, however, they might be associated with less abundant genera.

Culture pH interacts with corn oil concentration to affect biohydrogenation of unsaturated fatty acids and disappearance of neutral detergent fiber in batch culture

Effects of culture pH and corn oil (CO) concentration on biohydrogenation (BH) of unsaturated fatty acids and disappearance of neutral detergent fiber (NDF) in batch culture were evaluated in a 2 × 3 factorial design experiment. Culture vessels (100 mL; 4 replicates/treatment per time point) included ground alfalfa hay plus CO at 0, 1, or 2% dry matter inclusion rate and were incubated at pH 5.8 (low pH) or 6.2 (high pH) for 0, 6, 12, 18, or 24 h. Effects of culture pH, CO, time, and their interactions were determined. Adding CO increased total fatty acid concentration in substrates to 1.01, 2.31, and 3.58% dry matter for 0, 1, and 2% CO, respectively. Corn oil concentration interacted with culture pH and resulted in different effects on BH of cis-9,cis-12 18:2 at low or high culture pH. After 24 h of incubation, low pH, compared with high pH, reduced disappearance of NDF by 35% and BH extent of cis-9,cis-12 18:2 by 31%. Increasing CO increased disappearance of NDF across pH treatments and decreased BH extent of cis-9,cis-12 18:2 at low pH and increased it at high pH over 24 h. Compared with high pH, low pH reduced concentrations of 18:0 by 31% and increased concentrations of trans-10,cis-12 18:2 and trans-10 18:1 by 110 and 79% after 24 h, respectively. Adding CO at low pH had greater effect on BH intermediates of cis-9,cis-12 18:2 compared with adding oil at high pH. In particular, increasing CO to 1 and 2% DM at low pH, compared with at high pH, resulted in a 36 and 46% reduction in the concentration of 18:0, an 84 and 131% increase in the concentration of trans-10,cis-12 18:2, and an 81 and 129% increase in the concentration of trans-10 18:1, respectively. Despite the interactions between culture pH and CO concentration, main effects across time were also significant for the response variables of interest. In conclusion, culture pH interacted with CO concentration to affect BH of UFA and disappearance of NDF in batch culture, as the effects were greater at low culture pH than at high culture pH.

Rumen bacterial community responses to DPA, EPA and DHA in cattle and sheep: A comparative in vitro study

The role of marine lipids as modulators of ruminal biohydrogenation of dietary unsaturated fatty acids may be explained by the effects of their n-3 polyunsaturated fatty acids (PUFA) on the bacterial community. However, the impact of individual PUFA has barely been examined, and it is uncertain which bacteria are truly involved in biohydrogenation. In addition, despite interspecies differences in rumen bacterial composition, we are not aware of any direct comparison of bovine and ovine responses to dietary PUFA. Therefore, rumen fluid from cannulated cattle and sheep were used as inocula to examine in vitro the effect of 20:5n-3 (EPA), 22:5n-3 (DPA), and 22:6n-3 (DHA) on the bacterial community. Amplicon 16 S rRNA sequencing suggested that EPA and DHA had a greater contribution to the action of marine lipids than DPA both in cattle and sheep. Certain effects were exclusive to each ruminant species, which underlines the complexity of rumen microbial responses to dietary fatty acids. Based on changes in bacterial abundance, Barnesiella, Prevotella, Paraprevotella, Hallela, Anaerovorax, Succiniclasticum, Ruminococcus and Ruminobacter may be involved in the ruminal response in biohydrogenation to the addition of marine lipids, but further research is necessary to confirm their actual role in ruminal lipid metabolism.

The effect of dietary forage to concentrate ratio and forage type on milk fatty acid composition and milk fat globule size of lactating cows

We examined the effects of 2 grass silage-based diets differing in forage:concentrate (FC) ratio and those of a red clover silage-based diet on intake, milk production, ruminal fatty acid (FA) biohydrogenation, milk FA composition, and milk fat globule (MFG) size distribution. Ten multiparous Nordic Red cows received the following treatments: grass silage-based diets containing high (70:30, HG) or low (30:70, LG) FC ratio or a red clover silage-based diet with an FC ratio of 50:50 (RC) on a dry matter basis. Determinations of MFG were performed from fresh milk samples without addition of EDTA so the results of fat globules >1 µm in diameter are emphasized instead of the entire globule population. Lower FC ratio in grass silage-based diets increased milk production with no effect on daily fat yield, leading to 13% lower milk fat concentration. The effect of FC ratio on MFG size was moderate. It did not affect the volume-weighted diameter in grass silage-based diets, although LG lowered the volume-surface diameter of MFG in the size class >1 µm compared with HG. Compared with HG, feeding LG moderately decreased the biohydrogenation of 18:2n-6, leading to a higher level of polyunsaturated fatty acids in milk fat. Feeding RC lowered milk fat concentration and daily milk fat yield compared with grass silage-based diets. The volume-weighted diameter of MFG in the size class >1 µm was smaller in RC milk compared with grass silage-based diets. Feeding RC increased the flow of 18:3n-3 at the omasum by 2.4-fold and decreased the apparent ruminal 18:3n-3 biohydrogenation compared with grass silage-based diets despite similar intake of 18:3n-3. It also resulted in the lowest amount of saturated FA and the highest amounts of cis-9 18:1, 18:3n-3, and polyunsaturated FA in milk. In conclusion, LG decreased milk fat content and induced minor changes in MFG size distribution compared with HG, whereas RC lowered milk fat production, altered milk FA composition to nutritionally more beneficial direction, and led to smaller MFG compared with grass silage-based diets.

Physicochemical composition, fatty acid profile and sensory attributes of the meat of young Nellore bulls fed sunflower cake from the biodiesel industry

The use of agroindustrial waste to feed ruminants can change the microbial ecology of the animal, affect the fatty acid biohydrogenation extent and improve the profile of fatty acid of tissues. This study was conducted to determine the amount of sunflower cake in the diet of bulls that had the best effects on the physicochemical characteristics, fatty acid composition and sensory attributes of meat. To do so thirty-two intact Nellore bulls with an initial average body weight of 374 ± 42.5 kg that were used 24 months of age in a design completely randomized (four treatments). Animals were fed 400 g/kg Tifton 85 hay and 600 g/kg concentrate. The inclusion levels of sunflower cake (0, 9, 18, and 27% in DM basis) were the experimental treatments. Sunflower cake inclusion in the diet linearly increased the following characters (P <0.05): total lipid, protein, and cholesterol contents of meat; water-holding capacity; concentration of C18:0, C18:2n–6, ΣPUFA, Σn–6, and Σn–6:Σn–3 ratios; the h:H index; and the elongase enzymatic activity in the longissimus muscle of the bulls. However, the Warner-Bratzler shear force (WBSF), ash content, ΣUFA, ΣMUFA, ΣUFA:ΣSFA ratio and Δ9–desaturaseC16 enzymatic activity in the longissimus muscle of bulls showed a linear reduction (P <0.05) due to sunflower cake inclusion. A positive quadratic effect was observed for C18:2cis–9trans–11 (CLA) fatty acid, and a negative quadratic effect was observed for in the C18:3n–3. The level of inclusion of sunflower cake reduced the taster evaluation scores of the sensorial attributes regarding flavor, tenderness and overall acceptance compared to the control treatment (0%). Sunflower cake up to 21% of total DM could be fed to decrease feed costs without a major change in the fatty acid profile. However, the slight reduction in the sensory attributes and the increase in the most desirable fatty acids for health should be considered.

Consequences of inhibiting methanogenesis on the biohydrogenation of fatty acids in bovine ruminal digesta

Methanogenesis and the biohydrogenation of unsaturated fatty acids by ruminal microorganisms both utilize molecular (H2) or metabolic (2 H) hydrogen. The purpose of this study was to investigate the interdependence of the two metabolic activities, the first of which results in the emission of a potent greenhouse gas, the second causing ruminant products to have a low content of health-promoting unsaturated fatty acids. in vitro incubations were conducted using ruminal digesta taken post mortem from three beef cattle receiving a barley-based concentrate diet (C) and three grazing fresh grass (G). Inhibitors of methanogenesis, bromoethanesulfonate (BES), sodium nitrate and sodium nitrite, with or without added linoleic acid (LA; cis-9, cis-12-18:2), were added to determine the effects of inhibiting methanogenesis on biohydrogenation. These inhibitors have different modes of action and affect H2 metabolism differently. BS inhibits methanogenesis directly, and leads to the accumulation of H2, while nitrate and nitrite are alternative hydrogen sinks. BES inhibited 24-h CH4 formation by >96% and increased H2 accumulation with both diets; biohydrogenation and VFA production were unaffected, except for a small accumulation of trans-9-18:2 in G. Sodium nitrate inhibited methanogenesis by 82 and 58% with C and G, respectively, with no H2 accumulation; disappearance of LA was unaffected, though conjugated LA (CLA) isomers tended to accumulate in C but not G. Butyrate production was strongly inhibited by nitrate. Sodium nitrite inhibited accumulation of both H2 and CH4, but also volatile fatty acids, indicating that the overall fermentation was severely inhibited. qPCR of 16S rRNA genes indicated that members of the Butyrivibrio/Pseudobutyrivibrio clade increased in abundance during the incubation, while methanogens declined in C. It was concluded that inhibiting methanogenesis does not affect overall biohydrogenation, though nitrate may lead to lower butyrate production and increased CLA passage from the rumen.

Invited review: Plant polyphenols and rumen microbiota responsible for fatty acid biohydrogenation, fiber digestion, and methane emission: Experimental evidence and methodological approaches

The interest of the scientific community in the effects of plant polyphenols on animal nutrition is increasing. These compounds, in fact, are ubiquitous in the plant kingdom, especially in some spontaneous plants exploited as feeding resources alternative to cultivated crops and in several agro-industry by-products. Polyphenols interact with rumen microbiota, affecting carbohydrate fermentation, protein degradation, and lipid metabolism. Some of these aspects have been largely reviewed, especially for tannins; however, less information is available about the direct effect of polyphenols on the composition of rumen microbiota. In the present paper, we review the most recent literature about the effect of plant polyphenols on rumen microbiota responsible for unsaturated fatty acid biohydrogenation, fiber digestion, and methane production, taking into consideration the advances in microbiota analysis achieved in the last 10 yr. Key aspects, such as sample collection, sample storage, DNA extraction, and the main phylogenetic markers used in the reconstruction of microbial community structure, are examined. Furthermore, a summary of the new high-throughput methods based on next generation sequencing is reviewed. Several effects can be associated with dietary polyphenols. Polyphenols are able to depress or modulate the biohydrogenation of unsaturated fatty acids by a perturbation of ruminal microbiota composition. In particular, condensed tannins have an inhibitory effect on biohydrogenation, whereas hydrolyzable tannins seem to have a modulatory effect on biohydrogenation. With regard to fiber digestion, data from literature are quite consistent about a general depressive effect of polyphenols on gram-positive fibrolytic bacteria and ciliate protozoa, resulting in a reduction of volatile fatty acid production (mostly acetate molar production). Methane production is also usually reduced when tannins are included in the diet of ruminants, probably as a consequence of the inhibition of fiber digestion. However, some evidence suggests that hydrolyzable tannins may reduce methane emission by directly interacting with rumen microbiota without affecting fiber digestion.

Temporal changes in milk fatty acid composition during diet-induced milk fat depression in lactating cows

Diet-induced milk fat depression (MFD) in lactating cows has been attributed to alterations in ruminal lipid metabolism leading to the formation of specific fatty acid (FA) biohydrogenation intermediates that directly inhibit milk fat synthesis. However, the mechanisms responsible for decreased lipid synthesis in the mammary gland over time are not well defined. The aim of this study was to evaluate the effect of diet on milk FA composition and milk fat production over time, especially during MFD, and explore the associations between MFD and FA biohydrogenation intermediates in omasal digesta and milk. Four lactating Finnish Ayrshire cows used in a 4 × 4 Latin square with a 2 × 2 factorial arrangement of treatments and 35-d experimental periods were fed diets formulated to cause differences in ruminal and mammary lipid metabolism. Treatments consisted of an iso-nitrogenous total mixed ration based on grass silage with a forage to concentrate ratio of 65:35 or 35:65 without added oil, or with sunflower oil at 50 g/kg of diet dry matter. The high-concentrate diet with sunflower oil (HSO) induced a 2-stage drop in milk fat synthesis that was accompanied by specific temporal changes in the milk FA composition. The MFD on HSO was associated especially with trans-10 18:1 and also with trans-9,cis-11 conjugated linoleic acid (CLA) in milk and omasal digesta across all diets and was accompanied by the appearance of trans-10,cis-15 18:2. Trans-10,cis-12 CLA was increased in HSO, but milk fat secretion was not associated with omasal or milk trans-10,cis-12 CLA. The temporal changes in milk fat content and yield and milk FA composition reflect the shift from the predominant ruminal biohydrogenation pathway to an alternative pathway. The ambiguous role of trans-10,cis-12 CLA suggests that trans-10 18:1, trans-9,cis-11 CLA and trans-10,cis-15 18:2 or additional mechanisms contributed to the diet-induced MFD in lactating cows.