气相色谱测定羊瘤胃液中挥发性脂肪酸方法研究

建立了程序升温气相色谱法测定羊瘤胃液中的挥发性脂肪酸。样品经过滤离心,偏磷酸溶液酸化后,直接进样分析。结果表明:该方法实现了羊瘤胃液中乙酸、丙酸和正丁酸直接快速分离,方法的精密度、回收率较高。     

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.     

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.     

Supplementing Propionibacterium acidipropionici P169 does not affect methane production or volatile fatty acid profiles of different diets in in vitro rumen cultures from heifers

Abstract

Effects of Propionibacterium acidipropionici P169 on gas, methane (CH₄) production and volatile fatty acids in in vitro rumen cultures were studied. Ground barley (Experiment 1), mixed forage (Experiment 2) or mixed concentrate-forage (Experiment 3) served as substrates. Three doses of P169 were used in Experiments 1 and 2 [1.2 × 10⁴; 1.2 × 10⁶; 1.2 × 10⁸ colony forming unit (CFU)/mL], while two doses (2.4 × 10⁶; 2.4 × 10⁸ CFU/mL) were used in Experiment 3. In Experiment 1, P169 increased (P < 0.05) gas production at 6–12 h and CH₄ (mL/g diet dry matter) at all times. In Experiment 2, CH₄ was increased at 6 h (P < 0.01). At 12 h, propionate declined (P < 0.01) in Experiment 1 and increased at 48 h in Experiment 3. The inability of P169 to increase propionate and its lack of persistence in mixed ruminal cultures may account for its inability to alter CH₄ production.     

脂肪酸添加剂对奶牛瘤胃挥发性脂肪酸和甲烷的影响

21头安装瘤胃瘘管的荷斯坦产奶牛随机分为对照组和试验Ⅰ~Ⅵ组,每组3头,对照组饲喂全混日粮(TMR),试验Ⅰ~Ⅵ组每天饲喂TMR加50.0 g、100.0 g、150.0 g、200.0 g、250.0 g、300.0 g/头复合脂肪酸日粮,主要探讨复合脂肪酸对奶牛瘤胃甲烷(CH4)产量、挥发性脂肪酸(VFA)、氨氮(NH3-N)浓度和pH值及产奶性能的影响。结果,TMR中每天添加150.0~200.0 g/头复合脂肪酸,明显(P<0.05)提高奶牛乳脂率和瘤胃乙酸浓度,显著(P<0.05)降低奶牛的CH4释放量、CH4能/GE值和瘤胃NH3-N浓度,对奶牛的标准乳(FCM)、乳干物质、乳蛋白和乳糖均有提高趋势,能有效地提高奶牛的经济效益和生态效益。     

Degradation of two soluble proteins – casein and egg protein by a macro in vitro method

Degradation of casein and egg protein was studied with whole rumen contents (RC) in a macro in vitro system to elucidate previous findings of initial rapid disappearance of soluble proteins in vitro. Five to 7.5 kg of RC from a dry and/or a lactating cow were incubated with buffer and casein or egg protein for 180 min with frequent sampling. Degradation was measured as loss of trichloroacetic acid precipitable N (TCA‐N) from the inocula. Normal (39 °C) and low (2 °C) temperature incubations were examined in Exp. 1, using 1 g of TCA‐N from casein. Four levels of casein (0–12 g TCA‐N) in Exp. 2 and four levels of egg albumin (0–24 g TCA‐N) in Exp. 3 were fermented at 39 °C. Initial recovery of casein TCA‐N was 106% at 2 °C and 56% at 39 °C (Exp. 1). Casein (TCA‐N) recovered initially increased in Exp. 2 from 21% at 3 g to 86% at 12 g TCA‐N, while absolute loss remained relatively constant at 358 mg TCA‐N/kg RC (SD = 47). Fractional degradation rate was highest (0.03/min) at the intermediate dosage level. In the absence of rumen fluid (Exp. 4), no casein was lost. Initial egg protein recovery was on average 103% (Exp. 3). Recovery seemed unaffected by dosage level, and absolute degradation rate was relatively constant over time and increased with dosage level (p < 0.001) from 1.48 to 2.95 mg TCA‐N/(kg RC × min). Maximum degradation rate [mg TCA‐N/(kg RC × min)] and affinity constant (mg TCA‐N/kg RC) were estimated at 261 and 1650, respectively. It is concluded that a surprisingly constant amount of casein disappears immediately from warm rumen fluid and that this does not occur either with chilled RC, in the absence of rumen fluid, or when replaced with egg protein. The mechanisms for this disappearance are yet to be discovered.     

Effect of wheat processing on rumen characteristics and rumen parameters in Holstein‐Friesian calves

In this experiment, effect of wheat processing on rumen conditions and development were investigated. Fifty‐six neonatal Holstein‐Friesian calves (22 male and 34 female) were fed calf starters and post‐weaning diets containing 35 (pre‐weaning) and 21.90% (post‐weaning) popped wheat (PW), steam‐flaked wheat (SFW), dry‐rolled wheat (DRW) or ground wheat (GW) till 12 weeks of age. Calves were weaned at the end of 9th week, and a post‐weaning‐specific starter diets were fed for 1 month. Rumen liquor was analysed in days 30, 60 and 90 of the experiment to determine volatile fatty acids (VFA), pH and ammonia nitrogen concentrations. Twelve male calves (three calves/treatment) were slaughtered, and digestive tract was emptied. Forestomach empty weight and rumen parameters were assessed. Results indicated that calves received PW had the highest total VFA, acetate, propionate, butyrate, ammonia nitrogen, rumen wall thickness, papilla width and density. Calves fed DRW experienced the lowest rumen pH throughout the experiment probably because high proportion of fine particles in GW. Calves consuming PW apparently had more functional rumen in comparison with other groups.     

富含十八碳不饱和脂肪酸的植物油对体外瘤胃发酵和甲烷生成的影响

选取3种典型的植物油:菜籽油、豆油和亚麻油,利用体外培养系统进行体外发酵,研究植物油的类型和添加水平对瘤胃体外发酵和甲烷生成的影响。3种植物油分别添加底物干物质的3%、5%和7%,未添加植物油组为对照,共10个处理,每个处理设3个重复。结果表明,菜籽油对甲烷产量无显著影响(P>0.05),豆油7%添加水平显著抑制了甲烷生成,而亚麻油所有添加水平均显著降低了甲烷生成量(P<0.05)。在豆油和亚麻油处理组中,随着添加水平的提高,甲烷产量线性下降(P<0.01)。3种植物油均显著改变了瘤胃发酵模式,随着植物油中所含不饱和脂肪酸不饱和度的增加,乙酸和丁酸的摩尔比例显著降低,丙酸摩尔比例显著升高(P<0.05);植物油添加水平的提高显著降低了乙酸和丁酸摩尔比例,显著升高了丙酸摩尔比例(P<0.05),乙酸丙酸比例显著降低。本试验结果表明植物油降低甲烷的效果跟其来源和添加水平有关;随着植物油添加水平的提高和所含不饱和脂肪酸不饱和度的增加,甲烷生成量显著降低。     

瘤胃真菌体外培养条件对其产酶活性的影响

本文主要从环境因素、营养因素和其他因素等方面阐述了瘤胃真菌体外培养条件对其产酶活性的影响。     

瘤胃中乳酸的代谢及其调控机制

有关瘤胃酸中毒发生机制研究表明,瘤胃乳酸的累积可能对酸中毒诱导起重要作用,而高精料日粮下瘤胃乳酸累积主要取决于瘤胃乳酸产生菌和乳酸利用菌间的平衡程度。本文综述了瘤胃微生物对乳酸的代谢机制,包括主要乳酸产生菌[溶纤维丁酸弧菌(Butyrivibrio fibrisolvens)、牛链球菌(Streptococcus bovis)、乳酸杆菌(Lactobacillus)]和主要乳酸利用菌[反刍兽新月单胞菌(Selenomonus ruminantium)、埃氏巨型球菌(Megasphaera elsdenii)],并简要概述了酸中毒的调控方法,旨在为反刍动物瘤胃酸中毒的乳酸中毒机制深入解析提供参考。     

Rumen fermentation and degradability in buffalo and cattle using the in vitro gas production technique

An in vitro trial was conducted to investigate the effect of different inoculum sources (buffalo vs. cattle) on rumen fermentation and degradability. Incubations were carried out using rumen fluid obtained from buffalo or cattle fed the same diet [60% grass hay and 40% concentrate; 18 kg dry matter (DM)/day]. The fermentation kinetics of eight feeds commonly used in ruminant nutrition (alfalfa hay, barley meal, beet pulp, corn meal and silage, ryegrass hay and silage and soya bean meal s.e.) were studied with the in vitro gas production technique and rumen fermentation parameters (substrate disappearance, pH and volatile fatty acids production) were determined after 120 h of incubation. The linear relationship indicates that the microbial metabolic pathways of the two inocula for all the substrates were qualitatively similar, albeit often quantitatively different. In this in vitro study, a significant influence of rumen inoculum (buffalo vs. cow) on fermentation and degradability of the examined substrates was found. The differences in buffalo and cattle rumen fermentation can be explained with a different microbial activity of the two ruminant species, because of different amount of microbial population or microbial population constituted by different species of bacteria and protozoa.     

The microflora and concentrations of volatile fatty acids in the rumen of cattle fed on single component rations

The composition of the rumen microflora and the volatile fatty acids were examined in cattle free-grazing on grass or stall-fed on hay, grass pellets, oats or dried beet pulp with molasses. Total and viable counts of anaerobic bacteria were highest on the grass feeding, but viable counts as a percentage of total counts were highest when oats or beet pulp with molasses were fed. Counts of cellulolytic bacteria were lowest on these latter 2 diets, and highest on grass or grass pellet diets. Studies of the anaerobic flora showed that the composition in animals fed on grass pellets resembled more that found in animals free-grazing on grass than in those fed on hay. Counts of aerotolerant bacteria were only a small percentage of the total count, but were highest on the hay diet. On this latter diet and on grass-feeding the streptococci (identified as Streptococcus bovis) were predominant, but contrary to expectation, streptococci were found only in small numbers on the oats diet, where coryneform rods were the major type present. Although a period of 4–6 weeks was allowed for the animals to adapt to the feeds, the 2 periods of feeding on oats and dried beet pulp with molasses markedly affected the composition of the rumen flora in the subsequent periods of feeding grass pellets and hay. Ruinen volatile fatty acid analysis showed a propionogenic effect of oats and the highest percentage of butyric acid when beet pulp with molasses was fed. The expected propionogenic effect of grass pellets was not observed.     

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.     

Effects of supplementing an active dry yeast product on rumen microbial community composition and on subsequent rumen fermentation of lactating cows in the mid‐to‐late lactation period

The effects of supplementing feed of cows in mid‐to‐late lactation with an active yeast product (Actisaf Sc 47) were evaluated using 15 Holstein cows in a replicated 3 × 3 Latin square design. The animals were fed a mixed ration with 33% neutral detergent fiber, consisting of timothy hay (29.8%), a commercial concentrate (70.0%) and commercial calcium triphosphate (0.2%), twice daily to meet 105% of their energy requirement. Yeast supplement was set at 0, 5 and 10 g per day over 21‐day periods, each of which consisted of 14 days for adaptation followed by 7 days of data collection. Milking performance, plasma metabolite parameters, rumen volatile fatty acids, lipopolysaccharide and microbial properties were measured. Although there were no significant differences in feeding and milking performance or blood parameters associated with supplementation, the acetate to propionate ratio in the rumen fluid tended to decrease (P = 0.08). The population of Bacteroidetes tended to be less prominent (P = 0.07) and the fibrolytic bacterium Fibrobacter significantly increased (P < 0.05) in the rumen fluid of the yeast 10 g group compared with that of the control. These data suggest that effects of supplementing live yeast to cows in mid‐to‐late lactation may be limited to microbial composition and fermentation characteristics in the rumen.     

Effect of fatty acids added to the milk replacer on white scour and excretion of fatty acids in Holstein calves

In order to examine the relationship between white scour and fatty acids, we added fatty acids to the milk replacer. Twenty healthy Holstein calves were divided into 4 groups, five calves per group; a control group with no fortified fatty acid, and 3 groups fed either with oleic, stearic, or palmitic acid, respectively. The calves were fed milk replacer (5% of the calf's body weight) twice a day but the fatty acids (2 wt % of milk replacer) were added only once. The fecal and blood samples were obtained at 0, 12, 24, 36, and 48 h after feeding of the acids. All five calves in the palmitic acid group, and 3 out of 5 calves each in the stealic and the oleic acid groups had whitish feces after feeding fatty acid. The stearic acid group had a significantly elevated stearic acid concentration in the feces during 24–36 h compared to the pre-feeding level. The fecal concentration of palmitic acid increased significantly at 24–36 h in the palmitic acid group. We concluded that the long-chain saturated fatty acids are one of the causes of white scour in calves.     

Short-chain fatty acids inhibit bovine rumen epithelial cells proliferation via upregulation of cyclin-dependent kinase inhibitors 1A,but not mediated by G protein-coupled receptor 41

Short-chain fatty acids (SCFAs) play a critical role in regulation of rumen epithelial growth. The mechanisms underlying the regulatory effects of SCFAs on the proliferation of bovine rumen epithelial cells (BRECs) remain unknown; however, SCFAs can bind to G protein-coupled receptor 41 (GPR41); hence, the regulatory effects of SCFAs on BRECs proliferation may be mediated by GPR41. Here, we investigated the molecular mechanisms underlying the effects of SCFAs and GPR41 on BRECs proliferation. We demonstrated that SCFAs activate the expression of GPR41 and inhibit (p < .05) BRECs proliferation, while the GPR41 knockdown (GPR41KD) BRECs exhibited (p < .05) slow proliferation compared with controls. The treatment of BRECs with 10 mM SCFAs significantly enhanced (p < .05) expression of cyclin-dependent kinase inhibitors 1A (CDKN1A), 2A (CDKN2A) and 2B (CDKN2B) and inhibited (p < .05) their transition from G1 to S phase of the cell cycle, compared with controls. Remarkably, the GPR41KD BRECs treated with SCFAs restored high level of CDKN1A, relative to GPR41KD BRECs, but did not affect (p > .05) the expression of CDKN2A and CDKN2B. The GPR41KD BRECs had significantly reduced (p < .05) cyclin-dependent kinase 4 (CDK4) and cyclin D2 mRNA abundance compared with controls. The GPR41KD BRECs treated with SCFAs significantly decreased (p < .05) CDK4, cyclin D2, CDKN2A and CDKN2B mRNA abundance compared with BRECs treated with SCFAs. Overall, our results demonstrated that downregulation of CDK4 and cyclin D2 likely mediates the inhibitory effects of GPR41KD on BRECs proliferation. Additionally, CDKN1A plays a vital role in mediating the inhibitory effect of SCFAs on the BRECs proliferation, and that these changes are not mediated by GPR41.     

纳米铜对绵羊瘤胃体外发酵挥发性脂肪酸的影响

试验采用体外发酵法,研究在人工瘤胃液中添加不同浓度（0、50、100、150、200、400、800、1200、2400、4800μg/L）的纳米铜时,对绵羊瘤胃体外发酵挥发性脂肪酸的影响。试验共分10个处理组,每个处理组3个重复。结果表明：纳米铜在添加量为100-400μg/L时,乙酸浓度和总挥发性脂肪酸浓度都显著高于对照组和其他试验组（P〈0.05）。说明在体外发酵条件下添加适量的纳米铜对瘤胃微生物的发酵有促进作用。     

Effect of supplementation of rice bran and fumarate alone or in combination on in vitro rumen fermentation,methanogenesis and methanogens

This study investigated the effect of fumarate (FUM) and rice bran (RB), alone and together, on in vitro rumen fermentation, methanogenesis and methanogens. In vitro incubation was performed with six media that were either unsupplemented (control) or supplemented with 10% RB, 5 mmol/L FUM, 10% RB + 5 mmol/L FUM, 10 mmol/L FUM, or 10% RB + 10 mmol/L FUM. Methane (CH₄) production, dry matter digestibility, CH₄ per digested dry matter, total short‐chain fatty acid (SCFA) production, proportion of SCFA, acetate : proprionate ratio, production of NH₃‐N, and population density of rumen microbes were determined. Supplementation with 10% RB + 10 mmol/L FUM yielded a 36% decrease in CH₄ production compared to the control. Supplementation of FUM, in the presence or absence of RB, provided increases in total SCFA production and propionate proportion up to 61% and 31%, respectively. Total bacteria, methanogens and protozoa populations were significantly (P < 0.05) decreased with the 10% RB + 10 mmol/L FUM supplementation. The effect of anti‐methanogenesis of FUM was enhanced by the addition of RB. Notably, the CH₄ production attenuation was achieved by 10% RB + 10 mmol/L FUM without reduction of digestibility or of ruminal fermentation.     

不同种类异位酸添加水平对奶牛瘤胃体外发酵的影响

选用3头装有永久性瘤胃瘘管的奶牛,采用单因素对照试验设计和体外培养法研究不同种类异位酸的单独添加水平对奶牛瘤胃发酵的影响,并利用多项指标综合指数对培养液pH值、氨态氮浓度、菌体蛋白含量和残留有机物的含量进行综合评定,筛选3种异位酸单独使用的适宜添加量.结果表明,异丁酸0.2％DM和0.4％DM、异戊酸为0.15％DM和0.3％DM、2-甲基丁酸为0.1％DM和0.2％DM添加方式为最佳.     

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%。