Effects of medium‐chain fatty acid‐cyclodextrin complexes on ruminal methane production in vitro

The purpose of the present study was to evaluate effects of medium‐chain fatty acid‐cyclodextrin (CD) complexes on ruminal methane and volatile fatty acid production, and protozoal activity in vitro. Medium‐chain fatty acid‐CDs used in this study were caprylic acid (C8)‐αCD or ‐βCD, capric acid (C10)‐αCD or ‐βCD, and lauric acid (C12)‐αCD or ‐βCD. A 60‐mL of diluted rumen fluid was incubated anaerobically at 38°C for 6 h with the addition of the complex (10–40 mg as fatty acid). Each of the fatty acid‐CDs reduced the number of protozoa, with the order C10 > C12 > C8, and βCD complexes were more effective than αCD complexes. Molar proportions of acetic acid remained unchanged with the addition of fatty acid‐CD, while that of propionic acid increased, being significant for C8‐αCD and βCD, and C10‐αCD and βCD (P < 0.05). Hydrogen production decreased by about 70% of control with the addition of 40 mg of C8 and C10‐CD, on the other hand, it tended to increase with the addition of C12‐CD in both αCD and βCD. Methane production decreased by about 20% with the addition of 40 mg of complexes, except for C10‐βCD, which significantly reduced methane production by about 60%. In conclusion, the addition of C8 or C10‐CD to ruminant diets may be effective in reducing methane production.     

利用体外产气法研究2-甲基丁酸对山羊瘤胃发酵的影响

应用体外产气法模拟瘤胃发酵,通过测定24h体外累计产气量、细菌氮含量和中性洗涤纤维(NDF)的降解率,评价2-甲基丁酸的不同添加水平对山羊瘤胃发酵的影响。试验采用单因子设计,2-甲基丁酸的添加水平分别为0.0%、0.3%、0.6%、0.9%、1.2%,每个水平4个重复。结果发现:与对照组相比,添加0.3%的2-甲基丁酸组能极显著提高24h累计产气量(P<0.01);细菌氮含量有提高的趋势,但是差异不显著(P>0.05);在添加水平为0%～0.9%时,NDF降解率有提高的趋势,但是差异不显著(P>0.05),在1.2%时,其NDF降解率显著下降(P<0.05)。结果表明:日粮中的2-基丁酸的适宜添加水平为0.3%～0.6%。     

Quantitative studies of the in vitro production of pipecolic acid by rumen protozoa and its degradation by rumen bacteria

An in vitro study was conducted to quantitatively investigate the metabolism of pipecolic acid (Pip), a neuromodulator, by mixed rumen bacteria (B), mixed rumen protozoa (P), a combination of B and P (BP), species‐enriched rumen protozoal suspension (Polyplastron sp., Diploplastron sp., entodinia and Entodinium caudatum) and pure cultures of several isolates of rumen bacteria (Prevetolla bryantii, Prevetolla albensis, Streptococcus bovis, Veillonella parvula, Megasphaera elsdenii and Ruminococcus albus). Only P produced Pip from L‐lysine (1.0 mmol/L L‐Lys) at a rate of 83.5 ± 1.6 µmol/L/h and even in BP, Pip was produced from L‐Lys by P and increased at a rate of 31.2 ± 3.8 µmol/L/h. Pip production by P was highest when the substrate (L‐Lys) concentration was 6 mmol/L and then the rate was 580 ± 36 µmol/L/h. Pipecolic acid production by P suspension enriched with different species of protozoa showed that Polyplastron sp. had the highest Pip production rate of 0.907 ± 0.092 µmol/L/mg protozoal protein per h, and Diploplastron sp. had the lowest rate of 0.55 ± 0.13 µmol/L/mg protozoal protein per h. The addition of D‐Lys (1.0 mmol/L) as a substrate to the P suspension revealed that P were also able to produce Pip from D‐Lys, though at a lower rate (1/3) compared with L‐Lys (1.0 mmol/L), suggesting the presence of epimerases in P. It was confirmed that B were unable to produce Pip from L‐ or D‐Lys. Only B degraded Pip (1.0 mmol/L) after a lag phase at a rate of 56.0 ± 1.5 µmol/L/h. The B suspension was able to degrade D‐Lys, though the products were not identified. Pip degradation by pure culture of some species of rumen bacteria showed that P. bryantii and R. albus had the highest rate followed by P. albensis, S. bovis and M. elsdenii with a low rate of Pip degradation. Veillonella parvula showed no ability to degrade Pip. The results suggest that a fairly large proportion of rumen‐produced Pip is likely to be absorbed by the host animal before degradation by rumen bacteria.     

竹叶提取物对肉牛生产性能、瘤胃发酵特征及微生物数量的影响

试验研究竹叶提取物对肉牛生产性能、瘤胃发酵特征及微生物数量的影响.选取24头6月龄、体重(259.30±12.17) kg的西门塔尔杂交公牛,随机分为4组,每组6个重复.各组肉牛日粮中分别添加0(对照组)、40、80、120 g/d竹叶提取物,试验期180 d.结果 显示,80 g/d组肉牛的平均日增重和平均干物质采食...     

Fatty acid composition of ruminal bacteria and protozoa,and effect of defaunation on fatty acid profile in the rumen with special reference to conjugated linoleic acid in cattle

Objectives of this study were to compare fatty acid (FA) composition of ruminal bacterial (B) and protozoal (P) cells, and to investigate effect of protozoa on FA profile in the rumen of cattle. Three cows were used to prepare ruminal B and P cells. Four faunated and three defaunated cattle (half‐siblings) were used to study effect of protozoa on ruminal FA profile. Proportions of C16:0 and C18:0 in total fatty acids in B cells were 20.7% and 37.4%, whereas those in P cells were 33.4% and 11.6%, respectively. Proportions of trans‐vaccenic acid (VA) and cis‐9, trans‐11 conjugated linoleic acid (CLA) in B cells were 3.9% and 1.0%, and those in P cells were 5.5% and 1.6%, respectively, being higher in P cells. Proportions of C18:1, C18:2 and C18:3 in P cells were two to three times higher than in B cells. Proportions of unsaturated fatty acids, VA and CLA in B cells of faunated cattle were higher than those of defaunated. VA and CLA in the ruminal fluid of faunated were also 1.6 to 2.5 times higher than those of defaunated. This tendency was similar for cell‐free fraction of ruminal fluid. These results indicate that protozoa contribute greatly in VA and CLA production in the rumen.     

Effects of pure plant secondary metabolites on methane production,rumen fermentation and rumen bacteria populations in vitro

In this study, the effects of seven pure plant secondary metabolites (PSM s) on rumen fermentation, methane (CH₄) production and rumen bacterial community composition were determined. Two in vitro trials were conducted. In trial 1, nine concentrations of 8‐hydroxyquinoline, α‐ terpineol, camphor, bornyl acetate, α‐ pinene, thymoquinone and thymol were incubated on separate days using in vitro 24‐hr batch incubations. All compounds tested demonstrated the ability to alter rumen fermentation parameters and decrease CH₄ production. However, effective concentrations differed among individual PSM s. The lowest concentrations that reduced (p  <  .05) CH₄ production were as follows: 8 mg/L of 8‐hydroxyquinoline, 120 mg/L of thymoquinone, 240 mg/L of thymol and 480 mg/L of α‐ terpineol, camphor, bornyl acetate and α‐ pinene. These concentrations were selected for use in trial 2. In trial 2, PSM s were incubated in one run. Methane was decreased (p  <  .05) by all PSM s at selected concentrations. However, only 8‐hydroxyquinoline, bornyl acetate and thymoquinone decreased (p  <  .05) CH₄ relative to volatile fatty acids (VFA s). Based on denaturing gradient gel electrophoresis analysis, different PSM s changed the composition of bacterial communities to different extents. As revealed by Ion Torrent sequencing, the effects of PSM s on relative abundance were most pronounced in the predominant families, especially in Lachnospiraceae , Succinivibrionaceae , Prevotellaceae , unclassified Clostridiales and Ruminococcaceae . The CH ₄ production was correlated negatively (?.72; p  <  .05) with relative abundance of Succinivibrionaceae and positively with relative abundance of Ruminococcaceae (.86; p  <  .05). In summary, this study identified three pure PSM s (8hydroxyquinoline, bornyl acetate and thymoquinone) with potentially promising effects on rumen CH₄ production. The PSM s tested in this study demonstrated considerable impact on rumen bacterial communities even at the lowest concentrations that decreased CH₄ production. The findings from this study may help to elucidate how PSM s affect rumen bacterial fermentation.     

ORIGINAL ARTICLE: Description of development of rumen ecosystem by PCR assay in milk‐fed,weaned and finished lambs in an intensive fattening system

This study examined the reticulo‐rumen characteristics of the microbial community and its fermentative characteristics in milk‐fed, at weaning and finished lambs in a conventional fattening system. Five lambs were assigned to each of three groups: milk‐fed lambs slaughtered at 30 days (T30), weaned lambs slaughtered at 45 days (T45) and ‘finished lambs’ slaughtered at 90 days (T90). At slaughter, rumen size, fermentation parameters (pH, volatile fatty acids and microbial enzyme activity) and protozoal counts were recorded. Quantitative PCR was used to quantify the genes encoding 16S and 18S ribosomal DNA of the rumen bacterial and protozoal populations, respectively, and the sequential colonization of the rumen by cellulolytic (Ruminococcus albus, Ruminococcus flavefaciens) and amylolytic (Prevotella ruminicola, Streptococcus bovis) bacteria, and protozoa (Entodinium sp.). Denaturing gradient gel electrophoresis was used to study the development of rumen microbiota biodiversity. Intake of solid food before weaning caused a significant increase in rumen weight (p < 0.0001) and bacterial DNA (p < 0.05) and volatile fatty acid analysis concentration (p < 0.01), whereas pH declined. In milk‐fed lambs, cellulolytic bacteria were evident after 30 days. Thereafter, in the 45‐day and 90‐day groups, the proportions of R. flavefaciens decreased and R. albus increased. Amylolytic bacteria were present in milk‐fed lambs; the proportion of P. ruminicola increased in fattening lambs and S. bovis was the least abundant species. Protozoal concentrations were irregular; milk‐fed lambs had a significant number of protozoa species from Entodinium and subfamily Isotrichiidae, but they disappeared at weaning. Lamb rumen were refaunated in some individuals at 90 days (Entodinium and subfamily Diplodiniinae spp.), although individual concentrations were variable.     

大豆油不同添加水平对瘤胃体外发酵和产气的影响

本试验通过体外培养,研究在精粗比为30:70(偏粗型)日粮类型条件下,不同大豆油添加水平对产气量、甲烷产量、pH值和纤维降解率的影响.结果表明:随着大豆油添加水平的提高,产气量和甲烷产量逐渐降低,且各添加组显著低于对照组;pH值随着培养时间的延长,各添加水平在不同培养时间点与对照组相比均上升,其中添加量为4%和6%与对照组相比差异显著(P<0.05),添加量为8%时,与对照组相比差异极显著(P<0.01);培养24 h后.纤维降解率的抑制潜力8%水平最大,与对照组相比差异极显著(P<0.01),其次是6%的添加水平.与对照组相比差异显著(P<0.05).而2%和4%添加水平对纤维降解率的抑制程度与对照组相比差异不显著(P>0.05).通过产气量、pH值和纤维降解率综合考察大豆油对瘤胃体外发酵和产气的影响,本试验得出大豆油的最适添加量为4%.     

Gas and short‐chain fatty acid production from feeds commonly fed to red deer (Cervus elaphus L.) and incubated with rumen inoculum from red deer and sheep

Efficient red deer supplementary feeding depends on estimations of the nutritive value of offered feeds, frequently estimated with the use of equations derived from domestic ruminants. The aim of this study was to compare the 24‐hour in vitro true dry matter degradability (ivTD₂₄), in vitro gas production (GP) kinetic parameters, GP in 24 hr of incubation (GAS₂₄) and short‐chain fatty acid (SCFA) and microbial biomass (MBS) produced after 24‐hour incubation of feeds in inoculum prepared from sheep and red deer rumen fluid. Eleven feeds, frequently consumed by red deer in Slovenia, which occur either naturally (two fresh grasses, chestnut fruits and common and sessile oak acorns) or are fed as winter supplemental feeds (two grass hays, two grass silages, apple pomace, fresh sugar beetroot), were investigated. The in vitro GP kinetic parameters, GAS₂₄ and ivTD₂₄, did not differ between animal species. Amounts of SCFAs were greater (p < 0.05) when feeds were incubated in sheep inoculum, while molar proportions of acetic and propionic acids did not differ. Molar proportions of butyric acid produced during incubation of high fibre feeds did not differ between animal species, but were higher (p < 0.05) when feeds high in starch or sugar were incubated in red deer inoculum. Greater production of SCFA by sheep rumen microbes suggests better coverage of host animal with energy precursors, while greater production of MBS by red deer rumen microbes suggests better coverage of host animal with protein. Results also suggest that rumens of sheep and red deer are inhabited by different microbial communities, which did not affect the extent of in vitro GP and degradation of feeds used in the present experiment. However, the possibility exists that the divergent nutrient use could be a consequence of different priming by different feeds of the donor animal diets.     

Effects of rare earth element lanthanum on rumen methane and volatile fatty acid production and microbial flora in vitro

The objectives of the trial were to study the effects of rare earth element (REE) lanthanum (La) on the in vitro rumen methane (CH₄) and volatile fatty acid (VFA) production and the microbial flora of feeds. Four feed mixtures with different levels of neutral detergent fibre (NDF), that is 20.0% (I), 31.0% (II), 41.9% (III) and 52.7% (IV), were formulated as substrates. Five levels of LaCl₃, that is 0, 0.4, 0.6, 0.8 and 1.0 mmol/kg dry matter (DM), were added to the feed mixtures, respectively, as experimental treatments in a two‐factor 5 × 4 randomized design. The in vitro incubation lasted for 24 h. The results showed that supplementing LaCl₃ increased the total gas (p < 0.001) production and tended to increase the total VFA production (p = 0.072) and decreased the CH₄ production (p = 0.001) and the ratios of acetate/propionate (p = 0.019) and CH₄/total VFA (p < 0.001). Interactions between LaCl₃ and NDF were significant in total gas production (p = 0.030) and tended to be significant in CH₄ production (p = 0.071). Supplementing LaCl₃ at the level of 0.8 mmol/g DM decreased the relative abundance of methanogens and protozoa in the total bacterial 16S rDNA analysed using the real‐time PCR (p < 0.0001), increased F. succinogenes (p = 0.0003) and decreased R. flavefaciens (p < 0.0001) whereas did not affect R. albus and anaerobic fungi (p > 0.05). It was concluded that LaCl₃ decreased the CH₄ production without negatively affecting feed digestion through manipulating rumen microbial flora when feed mixtures with different levels of NDF were used as substrates.     

添加亚油酸条件下不同剂量硝酸钠对水牛瘤胃体外发酵脂肪酸组成及相关微生物数量的影响

为了探究添加亚油酸条件下不同剂量硝酸钠对水牛瘤胃体外发酵脂肪酸组成及相关微生物数量的影响,本试验选用3头装有永久性瘤胃瘘管的母水牛,以精粗比40:60为底物进行瘤胃液体外发酵试验.试验组硝酸钠的浓度分别为1、2、3 mg·mL-1,对照组不加硝酸钠,每组均添加0.25 mg·mL-1的亚油酸,每组各5个重复.在体外培养...     

日粮中添加不同水平红花油对绵羊瘤胃体外发酵动力学的影响

为探索红花油对瘤胃微生物发酵动力学的可能影响,试验以安装有永久性瘤胃瘘管的中国美利奴羊作为瘤胃微生物的供体动物,采用体外产气量法研究饲粮添加不同水平的红花油对绵羊瘤胃发酵动力学的影响。红花油的添加水平分别设0(对照组)、1%、2%和3%。结果表明:日粮添加2%和3%的红花油使产气速率和最大产气量明显低于对照组(P<0.05)。不同水平的红花油并未显著改变发酵液pH值、NH3-N浓度、CMCase及XYLase活性(P>0.05)。3%组的DM降解率略低于对照组(P<0.05),但与其他2个处理组差异不显著(P>0.05)。各处理组的NDF降解率较对照组也略有降低(P<0.05)。总挥发酸(TVFA)浓度、乙酸、丙酸摩尔百分比和乙丙酸比等指标各组间均差异不显著(P>0.05)。综上所述,饲粮中添加不超过3%水平的红花油对绵羊瘤胃发酵动力学没有负面影响。     

Theoretical turnover rate of the rumen liquid fraction in dairy cows and its relationship to feed intake,rumen fermentation,and milk production

Ruminant animals are able to convert plant materials (grain and the human‐indigestible portion of carbohydrates) to milk and meat. In this conversion, most of the plant materials are digested by rumen fermentation and are changed to short‐chain fatty acids, microbial cells, and methane, which is released into the atmosphere. The relationships among feed, rumen fermentation, and milk production are poorly understood. Here we report a novel indicator of characteristics of rumen fermentation, theoretical turnover rate (TTOR) of the rumen liquid fraction. The TTOR was calculated from the presumed rumen volume (PRV) which is estimated by dividing the methane yield by the methane concentration of rumen fluid. The formula for the TTOR is: TTOR = PRV/body weight^0.75. Our present analyses confirm that the TTOR as an indicator is capable of connecting feed, rumen fermentation, and milk production, because dry matter intake/TTOR showed a strong correlation with milk yield/TTOR. In addition, the TTOR may be related to ruminal pH, as we observed that the ruminal pH decreased as the TTOR increased. We propose that the TTOR is a factor characterizing rumen fermentation and a good indicator of the productivity of ruminants and dysbiosis of the rumen microbiome.     

不同油脂对瘤胃微生物体外产气及其区系动态变化的影响

本试验利用3头瘤胃瘘管山羊提供瘤胃液,以淀粉、纤维素、酪蛋白为底物进行体外培养,研究花生油、菜油、玉米油和豆油等对瘤胃发酵产气及微生物活力状况的影响。结果表明:培养液36 h总产气量在20.61～39.67mL,除花生油组与对照组差异不显著外(P>0.05),其他油脂组都显著低于对照组(P<0.05);随着培养时间的延长各组产气量呈现波动变化。培养液总脱氢酶以豆油组最高,显著高于对照组;并依次显著高于玉米油、花生油、菜籽油等组(P<0.05)。另外,培养液原虫DNA、细菌DNA、微生物DNA、原虫/细菌区系比例的均值与对照组,以及油脂组间差异都不显著(P>0.05),但随着培养时间的延长,微生物DNA在各时间点都以豆油与玉米油组的较高;原虫DNA一般在16 h达到最高,而细菌DNA则在8 h或16 h最高,并显著高于1 h或4 h的量(P<0.05、P<0.01),且各个试验组在培养过程中随时间的动态变化模式也不尽相同。总体看来,不同油脂对瘤胃微生物体外产气及区系动态变化的影响不同。     

Evaluation of sources of variation on in vitro fermentation kinetics of feedstuffs in a gas production system

The aim of this study was to evaluate the effect of different sources of variation in gas production technique on the in vitro gas production kinetics of feedstuffs. Triplicates of commercial concentrate, grass silage, grass hay and grass pasture were incubated in three experiments: experiment 1 assessed two agitation methods; experiment 2 evaluated different rumen inocula (pooled or different donor cows for each incubation run); and experiment 3 used Goering‐Van Soest or Mould buffers for media preparation. Gas production data were fitted into the Michaelis–Menten model and then subjected to analysis of variance. Gas production (GP) at 48 h and asymptote gas production (A) were lower when bottles were continuously under horizontal movement. Time to produce half and 75% of A, and A were affected by rumen inocula, while buffer type affected time to produce half and 25% of A and GP. No interactions between substrates and sources of variation were observed, suggesting that the effects of substrates on GP parameters were not modified. It is concluded that comparison of numerical data from in vitro experiments that follow different protocols must be done carefully. However, the ranking of different substrates is more robust and less affected by the sources of variation.     

瘤胃微生物体外发酵不同配比大豆荚与稻草混合粉产气量研究

为农村广泛开展稻草、玉米秸、麦秸、豆秸、谷草、荚壳等低粗饲料的有效利用,尤其是资源丰富的玉米秸秆和稻草,利用瘤胃微生物体外生物合成蛋白质饲料。试验设置7个处理组3重复,研究瘤胃微生物体外发酵大豆荚和稻草混合粉的产气量和气体种类。结果发现:大豆荚和稻草粉配比为7:3组合和6:4组合都能够促进羊瘤胃微生物体外繁殖与生长而提高饲料粗纤维的降解率。大豆荚和稻草混合粉配比7:3组合作为羊的瘤胃微生物体外发酵底物比较适宜。     

Effect of increasing levels of seven tree species extracts added to a high concentrate diet on in vitro rumen gas output

This study was conducted to investigate the effects of increasing levels of extracts of Byrsonima crassifolia, Celtis pallida, Enterolobium cyclocarpum, Fraxinus excelsior, Ficus trigonata, Phoradendrom brevifolium and Prunus domestica on in vitro gas production (GP) and ruminal fermentation of a high concentrate diet. Plant extracts were prepared at 1 g dry matter (DM)/8 mL of solvent mixture (methanol : ethanol : water, 1:1:8) and added at levels of 0, 0.6, 1.2 and 1.8 mL/g DM of a high concentrate diet. In vitro GP was recorded at 2, 4, 6, 8, 10, 12, 24, 48 and 72 h of incubation. Increasing addition of extracts linearly increased (P < 0.001), the GP₂₄, GP₄₈ and GP₇₂ (mL/g DM), and linearly decreased (P < 0.001), the discrete GP lag time. Moreover, increasing extract doses linearly increased (P < 0.001) the asymptotic GP and decreased (P < 0.001) the rate of GP. GP₆ was not impacted by treatments and GP₁₂ increased linearly (P = 0.01) with increasing addition of extracts. Rumen pH declined linearly (P < 0.05) with increasing doses of extracts added. As no interactions (P > 0.05) occurred between the extracts and doses, it could be conclude that all extracts positively modified rumen fermentation at doses of 1.2 to 1.8 mL extract/g diet DM.     

不同厌氧处理方式对鲜麦秸品质、活体外瘤胃发酵及甲烷产量的影响

为研究不同厌氧处理方式对鲜麦秸品质、活体外瘤胃发酵及甲烷产量的影响,试验分为5个处理组,分别为全株微贮组、秸秆黄贮组、秸秆微贮组、秸秆氨化组和秸秆碱化组。其中全株微贮组、秸秆微贮组秸秆经铡切处理后分别喷洒5×104cfu/g混菌悬浮液,秸秆黄贮组喷洒相同质量的无菌水;秸秆氨化组按照干物质5%喷洒尿素溶液,秸秆碱化组按秸秆干物质4%喷洒生石灰熟化澄清液。结果表明:秸秆微贮组、全株微贮组感官性能优于其他处理组;与秸秆黄贮组相比,全株微贮组、秸秆微贮组、秸秆氨化组及秸秆碱化组粗蛋白质含量分别提高219.21%(P<0.05)、16.75%(P>0.05)、112.81%(P<0.05)、13.79%(P>0.05);全株微贮组的中性洗涤纤维、酸性洗涤纤维含量约是其他处理组的1/2(P<0.05);与秸秆黄贮组相比,秸秆微贮组、秸秆氨化组、秸秆碱化组中性洗涤纤维显著降低8.24%、13.01%、9.09%,酸性洗涤纤维显著降低17.67%、17.68%及15.61%;产气量:全株微贮组>秸秆氨化组>其他处理组;秸秆黄贮组比全株微贮组、秸秆氨化组体外消化率降低了56.39%、24.03%;与秸秆微贮组相比,全株微贮组体外发酵液NH3-N、TVFA、乙酸、丙酸、丁酸、戊酸、异丁酸及异戊酸含量分别提高41.29%、20.99%、10.48%、41.99%、14.61%、166.67%、43.75%及64.71%(P<0.05)。由此可知,秸秆氨化处理和全株小麦经微生物处理均可提高其感官性能,提升产气量、挥发酸及消化率,全株小麦经微生物处理性能优于各秸秆处理组。     

Influence of Carotino oil on in vitro rumen fermentation,metabolism and apparent biohydrogenation of fatty acids

The study appraised the effects of Carotino oil on in vitro rumen fermentation, gas production, metabolism and apparent biohydrogenation of oleic, linoleic and linolenic acids. Carotino oil was added to a basal diet (50% concentrate and 50% oil palm frond) at the rate of 0, 2, 4, 6 and 8% dry matter of the diet. Rumen inoculum was obtained from three fistulated Boer bucks and incubated with 200 mg of each treatment for 24 h at 39°C. Gas production, fermentation kinetics, in vitro organic matter digestibility (IVOMD), volatile fatty acids (VFA), in vitro dry matter digestibility (IVDMD), metabolizable energy and free fatty acids were determined. Carotino oil did not affect (P > 0.05) gas production, metabolizable energy, pH, IVOMD, IVDMD, methane, total and individual VFAs. However, Carotino oil decreased (P < 0.05) the biohydrogenation of linoleic and linolenic acids but enhanced (P < 0.05) the biohydrogenation of oleic acid. After 24 h incubation, the concentrations of stearic, palmitic, pentadecanoic, myristic, myristoleic and lauric acids decreased (P < 0.05) while the concentration of linolenic, linoleic, oleic and transvaccenic acids and conjugated linoleic acid (CLAc9t11) increased (P < 0.05) with increasing levels of Carotino oil. Carotino oil seems to enhance the accumulation of beneficial unsaturated fatty acids without disrupting rumen fermentation.