体外产气法研究牛至油对绵羊瘤胃发酵特性和甲烷产量的影响

本试验采用体外产气法研究在体外培养液中添加牛至油对绵羊瘤胃发酵特性和甲烷产量的影响。采用单因素完全随机试验设计,在培养液中分别添加0(对照)、100、200、300和400 mg/L牛至油,每组5个重复。发酵12 h后,测定总产气量、甲烷产量及培养液p H,总氮、氨氮、尿素氮、蛋白氮浓度及挥发性脂肪酸浓度和比例。结果表明:1)培养液中添加100、200、300和400 mg/L牛至油均显著升高了培养液p H(P0.05);高添加水平(300、400 mg/L)的牛至油显著降低了培养液总挥发性脂肪酸浓度和丙酸比例(P0.05),显著升高了培养液乙酸比例和乙酸/丙酸(P0.05),抑制了瘤胃发酵。2)与对照组相比,除100 mg/L牛至油组外,其余各试验组培养液氨氮浓度均显著降低(P0.05)。3)高添加水平(300、400 mg/L)的牛至油在显著降低甲烷产量的同时也显著降低了总产气量(P0.05)。由此可知,牛至油能调控瘤胃发酵和降低甲烷产量,但过高添加水平对绵羊瘤胃发酵有抑制作用,体外条件下最适添加水平为200 mg/L。     

不同植物精油对体外瘤胃发酵及甲烷产量影响的比较研究

在底物精粗比为6∶4的条件下,在底物中添加不同剂量[使发酵液中植物精油的浓度分别为0(对照)、50、100、200和400 mg/L]的丁子香酚、D-柠烯、茴香脑、肉桂醛、百里香酚或香芹酚,通过体外产气法比较研究不同植物精油对体外瘤胃发酵和甲烷(CH4)产量的影响。每种植物精油的每个剂量设3个重复。体外模拟瘤胃发酵培养24 h,测定产气量和气体中的CH4含量以及发酵液的p H、挥发性脂肪酸(VFA)和氨态氮(NH3-N)浓度。结果表明:1)除百里香酚外,添加各种植物精油对体外发酵液p H均无显著影响(P0.05)。2)添加丁子香酚、D-柠烯、茴香脑和肉桂醛对体外发酵液总VFA浓度没有显著影响(P0.05),但总VFA浓度随百里香酚和香芹酚浓度的增加呈二次曲线变化(PQ0.01)。与对照组相比,添加400 mg/L百里香酚和香芹酚显著降低体外发酵液总VFA浓度(P0.01)。D-柠烯、茴香脑、百里香酚和香芹酚的添加改变了各VFA占总VFA的摩尔百分比。与对照组相比,添加50 mg/L D-柠烯和茴香脑使乙酸比例显著增加(P0.05),丙酸比例显著降低(P0.05);而添加400 mg/L D-柠烯和茴香脑则使乙酸比例显著下降(P0.05),丙酸和丁酸比例显著上升(P0.05)。百里香酚和香芹酚的添加对乙酸比例没有产生显著影响(P0.05),与对照组相比,400 mg/L百里香酚和香芹酚使丙酸比例显著下降(P0.05)。3)添加茴香脑、百里香酚和香芹酚显著影响体外发酵液NH3-N浓度(P0.05),与对照组相比,400 mg/L百里香酚和香芹酚显著降低NH3-N浓度(P0.05)。4)添加D-柠烯、茴香脑、肉桂醛对体外发酵24 h产气量没有显著影响(P0.05)。与对照组相比,各浓度的百里香酚和香芹酚均显著降低体外发酵24 h产气量(P0.05),且产气量随百里香酚和香芹酚浓度的增加呈二次曲线变化(PQ0.01)。5)添加D-柠烯、茴香脑和肉桂醛对体外发酵24 h CH4产量没有显著影响(P0.05)。与对照组相比,50和100 mg/L的丁子香酚显著增加体外发酵24 h CH4产量(P0.05),而400 mg/L的百里香酚和香芹酚体外发酵24 h CH4产量分别降低84.7%(P0.05)和73.9%(P0.05)。综合以上试验结果可知,不同植物精油对体外瘤胃发酵和CH4产量的影响结果不同,且与添加剂量有关。其中,低剂量的百里香酚和香芹酚促进体外瘤胃发酵,而高剂量的百里香酚和香芹酚抑制体外瘤胃发酵且显著降低24 h CH4产量。     

利用双外流持续发酵系统研究植物精油对瘤胃发酵和甲烷生成的影响

本研究采用双外流持续培养法,探讨添加不同水平的植物精油对瘤胃发酵和甲烷(CH4)生成的影响。每种植物精油(肉桂油和留兰香油)的4种添加水平(0、100、500和1 500 mg/L)作为处理因素,每个处理3个重复,每个试验周期为6 d。结果表明:与对照组相比,1)添加1 500 mg/L的肉桂油显著降低了发酵液pH、饲粮中性洗涤纤维降解率(dNDF)、酸性洗涤纤维降解率(dADF)和粗蛋白质降解率(dCP)(P<0.05)。2)添加100 mg/L的留兰香油和肉桂油对挥发性脂肪酸(VFA)浓度和乙酸/丙酸(A/P)无显著影响(P>0.05)。添加500 mg/L的肉桂油在发酵27和99 h以及添加1 500 mg/L的肉桂油在所有发酵时间点均显著降低了总挥发性脂肪酸(TVFA)、乙酸和丙酸浓度(P<0.05)。随着发酵时间的延长,1 500 mg/L肉桂油添加组TVFA、乙酸和丙酸浓度显著下降(P<0.05),A/P显著升高(P<0.05)。3)添加500 mg/L的肉桂油以及1 500 mg/L的肉桂油和留兰香油均显著降低了发酵液中的原虫数量(P<0.05)。添加3种剂量的肉桂油和留兰香油均显著降低了CH4体积分数(P<0.05)。由此可见,添加100 mg/L的肉桂油和留兰香油均有助于改善瘤胃微生物发酵。     

葡萄籽精油对体外瘤胃发酵和甲烷生成的影响

通过体外培养法研究发酵底物精粗比7∶3条件下,添加100,150和250μL/L葡萄籽精油对瘤胃发酵和甲烷产量的影响。采用气相色谱仪测定甲烷产量,培养24 h后,测定挥发性脂肪酸、pH、氨态氮浓度和原虫数。结果发现,添加葡萄籽精油100μL/L和150μL/L与对照组相比提高累计产气量、发酵液pH值(P<0.05)、乙酸摩尔比例,降低甲烷浓度(P<0.01)、丙酸、丁酸、戊酸和支链挥发性脂肪酸摩尔比例。葡萄籽精油添加量为150μL/L降低氨态氮浓度及瘤胃原虫数,添加100μL/L葡萄籽精油提高总挥发性脂肪酸。试验表明,适宜添加量的葡萄籽精油可以改变瘤胃发酵模式,增加丙酸乙酸比例,抑制甲烷生成。     

活性干酵母和纤维素酶对草原红牛瘤胃挥发性脂肪酸浓度及甲烷排放的影响

利用大型动物开放式呼吸测热装置等设备,研究活性干酵母和纤维素酶对草原红牛瘤胃挥发性脂肪酸浓度及甲烷排放的影响。试验选用8月龄、体况相近的草原红牛公牛4头,采用4×4拉丁方试验设计分为4组:Ⅰ组为对照组,饲喂基础日粮,Ⅱ组在基础日粮中添加200 mg/kg活性干酵母,Ⅲ组在基础日粮中添加190 mg/kg纤维素酶,Ⅳ组在基础日粮中添加200 mg/kg活性干酵母和190 mg/kg纤维素酶,试验分4期,每期37 d。结果表明,日粮中添加活性干酵母后,对瘤胃内丙酸浓度和总挥发性脂肪酸浓度影响显著(P<0.05),Ⅱ、Ⅳ组总挥发性脂肪酸浓度和丙酸浓度显著高于Ⅰ、Ⅲ组(P<0.05);活性干酵母显著抑制了草原红牛甲烷的产生(P<0.05),Ⅱ组甲烷排放量比Ⅰ组降低9.84%;纤维素酶对草原红牛瘤胃挥发性脂肪酸及甲烷排放的影响均不显著(P>0.05)。因此,日粮中添加活性干酵母能促进草原红牛瘤胃发酵,增加丙酸浓度,并显著降低甲烷排放,减少能量损失。     

体外产气法研究植物精油对肉羊体外瘤胃发酵参数及甲烷产量的影响

本试验旨在采用体外产气法研究4种天然植物精油(桉叶油、山苍子油、肉桂油和茴香油)对肉羊体外瘤胃发酵参数及甲烷(CH4)产量的影响。体外培养底物精粗比为60∶40,分别添加0(对照)、50、100、200和400 mg/L的桉叶油、山苍子油、肉桂油和茴香油,每种植物精油的每个浓度设置3个重复,体外模拟瘤胃发酵培养24 h,测定24 h产气量和气体中的CH4产量以及瘤胃发酵液的pH、挥发性脂肪酸(VFA)和氨态氮(NH3-N)浓度。结果表明:1)添加山苍子油、茴香油显著影响了体外瘤胃发酵液pH(P0.05)。2)与对照组相比,添加400 mg/L山苍子油、茴香油显著降低了总VFA浓度(P0.05),且随着山苍子油、茴香油添加浓度的增加呈线性下降趋势(P0.05);添加400 mg/L山苍子油和茴香油均显著增加了乙酸比例(P0.05),且随着山苍子油和茴香油添加浓度的增加呈先降低后升高的二次曲线趋势(P0.05);添加200 mg/L茴香油、400 mg/L桉叶油显著提高了丙酸比例(P0.05),而添加400 mg/L的山苍子油显著降低了丙酸比例(P0.05),且随着桉叶油、山苍子油、茴香油添加浓度的增加呈二次曲线变化趋势(P0.05);添加400 mg/L桉叶油、200 mg/L茴香油显著降低了乙酸/丙酸(P0.05),添加400 mg/L山苍子油显著增加了乙酸/丙酸(P0.05),且随着桉叶油、山苍子油、茴香油添加浓度的增加呈二次曲线变化趋势(P0.05)。3)与对照组相比,添加400 mg/L茴香油显著降低了NH3-N含量(P0.05),且随着茴香油添加浓度的增加呈线性下降趋势(P0.05)。4)与对照组相比,添加400 mg/L山苍子油、茴香油均显著降低了产气量(P0.05),且随则山苍子油添加浓度的增加呈先增加后降低的二次曲线趋势(P0.05),随着茴香油添加浓度的增加呈线性下降趋势(P0.05);添加400 mg/L山苍子油、茴香油显著降低了CH4产量(P0.05),且随着山苍子油添加浓度的增加呈先增加后降低的二次曲线趋势(P0.05),随着茴香油添加浓度的增加呈线性下降趋势(P0.05)。由此可见,不同植物精油对体外瘤胃发酵参数和CH4产量的影响结果不同,且与植物精油浓度有关。     

硝酸钠和2-溴乙烷磺酸钠对山羊体外瘤胃发酵甲烷、氢气和挥发性脂肪酸生成的影响

本试验从瘤胃氢代谢生物化学过程的角度出发,研究硝酸钠和2-溴乙烷磺酸钠(BES)对瘤胃甲烷生成的抑制机制。选用2只装有永久性瘤胃瘘管的成年湘东黑山羊作为瘤胃液供体,设对照组、硝酸钠组(添加剂量分别为0.10、0.20和0.40 mg/m L)和BES组(添加剂量分别为0.63、1.26和2.52 mg/L),采用全自动体外模拟瘤胃发酵设备,进行24 h体外瘤胃发酵试验,测定产气量、氢气和甲烷的产量和含量、挥发性脂肪酸产量及组成。结果表明:与对照组相比,添加硝酸钠和BES后,甲烷产量显著降低(P0.05),并且随添加剂量增加,呈现线性下降的变化趋势(P0.05)。低剂量硝酸钠(≤0.20 mg/m L)对产气量、起始底物降解速率、乙酸、丙酸产量和乙丙比无显著影响(P0.05);高剂量硝酸钠(0.40 mg/m L)显著提高了氢气产量及含量,显著降低了产气量、甲烷产量及含量、起始底物降解速率和总挥发性脂肪酸产量(P0.05);添加BES对产气量、起始底物降解速率和总挥发性脂肪酸产量没有显著影响(P0.05),但显著增加了氢气产量及含量(P0.05),显著降低了乙丙比(P0.05),发酵类型向丙酸型转变。由此可见,硝酸钠减少甲烷生成依靠其氢池和瘤胃微生物毒性作用,而BES减少甲烷生成依靠抑制甲烷产生,对气体生成和饲料的降解影响很小。     

体外产气法评价植物提取物对生长肥育猪粪样微生物发酵特性及甲烷生成的影响北大核心CSCD

本试验旨在通过体外产气法研究牛至油、肉桂油及大蒜素3种植物提取物对生长肥育猪粪样微生物发酵特性及甲烷(CH_(4))生成的影响。采用Menke体外产气法,在含饲粮底物和生长肥育猪鲜粪滤液的产气瓶中分别添加0(对照)、200、400和800 mg/L的牛至油、肉桂油或大蒜素,在39℃下培养48 h,测定不同时间点(0、2、6、12、24和48 h)的产气量、CH_(4)产量、产气动力学参数及短链脂肪酸(SCFA)浓度。结果表明:1)与对照组相比,3种植物提取物均显著降低了48 h累积产气量(P<0.05),其中800 mg/L牛至油组下降幅度最大。2)与对照组相比,400 mg/L牛至油组和800 mg/L肉桂油组产气动力学参数显著降低(P<0.05)。3)与对照组相比,除200 mg/L牛至油组外,各植物提取物组48 h累积CH_(4)产量均显著降低(P<0.05),且各800 mg/L植物提取物组发酵2 h后各时间点所产生的气体中CH_(4)浓度均显著降低(P<0.05)。4)200 mg/L植物提取物组乙酸和总SCFA浓度从高到低依次为大蒜素组>牛至油>肉桂油组;与对照组相比,各800 mg/L植物提取物组乙酸、丙酸和总SCFA浓度均显著降低(P<0.05),各大蒜素组丁酸浓度均显著提高(P<0.05),同时400 mg/L大蒜素组乙酸/丙酸值显著降低(P<0.05)。综上可知,不同植物提取物均会影响生长肥育猪粪样微生物发酵特性,降低体外发酵累积产气量和CH_(4)产量,其中各植物提取物浓度为800 mg/L时对体外发酵与CH_(4)生成的抑制效果最强,大蒜素浓度为200 mg/L时能够在投入更低的成本抑制CH_(4)产生的同时表现出更好的发酵特性。     

不同植物精油及精油成分体外抗氧化性能评定

为比较天然植物精油与精油活性成分单体的抗氧化性能，采用清除1,1-二苯基-2-苦基肼（1, 1-Diphenyl-2-Picrylhydrazyl, DPPH）自由基的方法，以抗氧化剂2,6-二叔丁基-4-甲基苯酚(抗氧化剂，BHT)为对照，测定了牛至油、丁香油、薰衣草油等5种植物精油和丁香酚、香芹酚、肉桂醛等20种精油成分的体外抗氧化能力，从中筛选出DPPH自由基清除率在90%以上植物精油和精油成分，以及清除率低于90%，但生产中常用的柠檬醛、肉桂醛、柠檬烯，测定其IC50值。结果表明：(1)在浓度为200 mg/mL时，牛至油、丁香油、薰衣草油、肉桂油、缬草油、丁香酚、百里香酚、香芹酚、松油醇、香草醛和BHT对DPPH自由基的清除率在90%以上；柠檬醛、肉桂醛和柠檬烯的清除率分别是84.79%、45.55%、36.21%。(2)当浓度降为20 mg/mL时，牛至油、丁香油、薰衣草油、丁香酚、百里香酚、香芹酚、香草醛和BHT对自由基的清除率仍在90%以上，但肉桂油、缬草油和松油醇的清除率有所降低，分别为48.90%、38.51%、43.01%。(3)根据IC50值，几种精油和精油成分...     

3种饲用植物精油及其主要成分对猪源致病菌的抑菌作用研究

采用高效液相色谱法,检测3种可饲精油(肉桂油、牛至油和百里香油)中主要成分的含量,用微板指示剂法测定3种精油及其主要成分对猪源产肠毒素大肠杆菌K88和霍乱沙门菌A72的体外抑菌活力。结果表明:肉桂油及其主要成分的抑菌活力最强,最小抑菌质量浓度(MIC)为400~800μL/L;百里香油及其主要成分百里香酚的作用次之,MIC为1 600μL/L;牛至油及其主要成分香芹酚的作用最差,MIC为1 600~3 200μL/L。     

Effects of the essential oil cyclodextrin complexes on ruminal methane production in vitro

The effects of including essential oil cyclodextrin (CD) complexes on ruminal methane and hydrogen production, volatile fatty acid (VFA) and protozoa were studied in vitro. The essential oil CD complexes used in the present study were both αCD and βCD with cineol, eucalyptus, menthol, peppermint, thyme and wasabi. Diluted rumen fluid (60 mL) was incubated anaerobically at 38°C for 6 h with the essential oil CD complex (1–40 mg as oil). Eucalyptus‐αCD reduced methane production by 40% with 10 mg as oil along with decreasing the protozoal number and increasing total VFA and proportion of propionic acid. Both wasabi‐αCD and wasabi‐βCD reduced methane production by 85% and 97%, respectively, with 10 mg as oil, though they increased hydrogen production notably. The protozoal number was unchanged by wasabi‐βCD and decreased to 50% by wasabi‐αCD. Propionic acid was increased by both wasabi‐αCD and wasabi‐βCD. The other essential oil‐CD showed no significant effect on reducing methane production. The combined use of 20 mM of fumaric acid with wasabi‐αCD (1 mg as oil) caused about a 50% depression of methane production compared to the control without increasing hydrogen production. Fumaric acid was more effective than malic acid in reducing methane production. The addition of eucalyptus‐αCD, wasabi‐αCD or wasabi‐βCD together with organic acid to rumen fluid turned out to be effective for reducing methane production with no increase in hydrogen production.     

添加不同水平单宁酸对绵羊体外发酵参数和甲烷产量的影响

本试验以精料、青干草和单宁酸作为发酵底物,研究添加不同水平单宁酸对绵羊体外发酵参数和甲烷产量、干物质和粗蛋白质降解率的影响,以期为体内试验筛选适宜的单宁酸浓度,为富含单宁的粗饲料资源应用于反刍动物日粮配制及减少反刍动物养殖过程种甲烷排放提供科学依据。采用体外批次培养法开展研究,发酵底物为精料、青干草和单宁酸,设置底物的精粗比为3∶7。根据单宁酸添加量分为6组,单宁酸添加量分别为0、0.5%、1.0%、1.5%、2.0%、3.0%,发酵时间设置为3、6、12和24 h,每个处理3个重复,测定各培养时间pH、氨氮（NH₃-N）浓度、挥发性脂肪酸（VFA）浓度、总产气量、甲烷产量、干物质和粗蛋白质降解率。结果显示,培养至3 h时,1.0%和2.0%单宁酸添加组pH显著高于对照组（P<0.05）,其他时间点各组pH无显著差异（P>0.05）;1.0%、2.0%和3.0%单宁酸添加组NH₃-N浓度显著低于对照组（P<0.05）;培养至6和24 h时,各处理组NH₃-N浓度均显著低于对照组（P<0.05）。培养至6 h时,2.0%单宁酸添加组乙酸、丙酸、丁酸和总VFA浓度均显著低于对照组（P<0.05）;培养至24 h时,1.0%、1.5%和3.0%单宁酸添加组的丙酸、丁酸和总VFA浓度均显著低于对照组（P<0.05）;各单宁酸添加组4个时间点丙酸、丁酸、总VFA浓度的平均值均显著低于对照组（P<0.05）。培养至12、24 h时,各处理组总产气量与对照组差异不显著（P>0.05）;培养至12 h时,2.0%组甲烷产量显著低于对照组、0.5%组和1.0%组（P<0.05）;培养至24 h时,2.0%和3.0%组的甲烷产量显著低于对照组（P<0.05）。因此,添加单宁酸会显著降低体外发酵的NH₃-N浓度,抑制VFA的产量,添加1.5%、2.0%的单宁酸能够显著降低瘤胃发酵甲烷产量,添加单宁酸的处理组粗蛋白质降解率显著低于对照组。     

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.     

In vitro gas formation and fermentation parameters using different substrates and pig faecal inocula affected by bile extract

The effect of bile extract on microbial gas formation and composition, production of volatile fatty acids (VFA) and methane and on reductive acetogenesis was studied in vitro. As substrates cellulose and potato starch in combination with graded levels of bile extract concentrations (0; 333; 666; 1333 mg/l) were incubated with a faecal suspension from pigs as microbial inoculum according to procedures of a modified “Hohenheim gas formation test” (HFT). Bile extract concentration of 1333 mg/l in the in vitro system reduced total gas production from cellulose significantly (p < 0.05) compared to incubation without bile extract. After 24 h of incubation, bile extract concentration of 1333 mg/l lead to a decrease in methane production from cellulose (p < 0.05) in comparison to incubation without bile extract. Increasing levels of bile extract reduced total production of VFA (p < 0.05) from cellulose. Furthermore, bile extract concentration of 1333 mg/l increased molar percentage of propionate from cellulose and starch after 24 h of incubation (p < 0.05). Bile extract concentration of 333 mg/l increased molar percentage of propionate from cellulose after 48 h of incubation (p < 0.05). The results may contribute to explain the differences in gas and methane production between the rumen and the hindgut of monogastric animals.     

用体外发酵法评价亚热带几种常用反刍动物饲料的营养价值

试验旨在用体外发酵法评价亚热带多种常用反刍动物饲料的营养价值。选用甘蔗尾、紫色象草、桂闽引象草、构树、玉米秸秆、菠萝皮、鲜豆腐渣、鲜啤酒渣和罗汉果渣9种饲料作为发酵底物,通过瘘管采集成年湘东黑山羊的瘤胃液,利用全自动体外模拟瘤胃发酵设备进行48 h体外发酵。根据总产气量（TGP）、甲烷产量、干物质降解率（DMD）以及挥发性脂肪酸（VFA）浓度等发酵参数进行评价。结果表明：几种料渣中,48 h总产气量、甲烷产量、干物质降解率和VFA浓度由高到低的顺序均为鲜豆腐渣、菠萝皮、鲜啤酒渣、罗汉果渣;几种草料中,玉米秸秆的甲烷产量最高,构树的48 h总产气量、干物质降解率和VFA浓度最高,罗汉果渣的这几项指标均最低。由上述结果可知,从常规营养成分、体外发酵产气和挥发性脂肪酸浓度等方面综合考虑,除罗汉果渣外,其他8种常用饲料均可作为反刍动物的优质饲料加以开发利用。     

Rumen microbial response in production of CLA and methane to safflower oil in association with fish oil or/and fumarate

Supplementation effect of fish oil and/or fumarate on production of conjugated linoleic acid (CLA) and methane by rumen microbes was examined when incubated with safflower oil. One hundred and twenty milligrams of safflower oil (SO), safflower oil with 24 mg fish oil (SOFO), safflower oil with 24 mmol/L fumarate (SOFA), or safflower oil with 24 mg fish oil and 24 mmol/L fumarate (SOFOFA) were added to the 90 mL culture solution. The culture solution was also made without any supplements (control). The SOFA and SOFOFA increased pH and propionate (C3) compared to other treatments from 3 h incubation time. An accumulated amount of total methane (CH₄) for 12 h incubation was decreased by all the supplements compared to control. The concentrations of c9,t11CLA for all the incubation times were increased in the treatments of SOFO, SOFA and SOFOFA compared to SO. The highest concentration of c9,t11CLA was observed from SOFOFA among all the treatments at all incubation times. Overall data indicate that supplementation of combined fumarate and/or fish oil when incubated with safflower oil could depress CH₄ generation and increase production of C₃ and CLA under the condition of current in vitro study.     

In vitro rumen fermentation and methane production are influenced by active components of essential oils combined with fumarate

Two trials were conducted to identify the optimal levels of essential oil active components (EOAC) and their combination with fumarate on in vitro rumen fermentation. In trial 1, eugenol, carvacrol, citral and cinnamaldehyde were mixed at ratios of 1:2:3:4, 2:1:4:3, 3:4:1:2, 4:3:2:1 and 1:1:1:1 to make up five combinations (EOAC1, EOAC2, EOAC3, EOAC4 and EOAC5 respectively). The mixtures were supplied at levels of 0, 50, 200 or 500 mg/l to identify the optimal combination for methane reduction. Methane production and ammonia nitrogen were decreased by adding EOAC, irrespective of component compounds, but the production of gas and total volatile fatty acids (VFA) were also decreased. Hydrogen balance analysis indicated that the ratio of hydrogen consumed via methane to hydrogen consumed via VFA was lowest at 200 mg/l of EOAC5 treatment, from which the proportional change in methane was more than the change in VFA, with 31.5% of methane reduction and 12.9% of VFA reduction. In trial 2, 200 mg/l of EOAC5 was added with 0, 5, 10 and 15 mm monosodium fumarate to see whether fumarate had a further effect on rumen fermentation. The addition of fumarate had no influence on gas production, but it further decreased methane and increased the total VFA in comparison with EOAC added solely, with the greatest decrease occurring in methane (78.1%) from 10 mm of fumarate. Quantification of the microbial populations in rumen fluids by RT‐PCR showed that methanogen, protozoa, fungi, Fibrobacter succinogenes and Ruminococcus flavefaciens populations were significantly decreased by EOAC5, but were not influenced by fumarate. In summary, the addition of EOAC had consistent effects on rumen fermentation parameters, but high levels of EOAC would induce the inhibition of rumen fermentation. Adding fumarate can enhance the methane‐inhibiting effect of EOAC, and the decrease was higher than that calculated stoichiometrically.     

Effect of traditional Chinese medicine compounds on rumen fermentation,methanogenesis and microbial flora in vitro

This study was conducted to investigate the effects of traditional Chinese medicine compounds (TCMC) on rumen fermentation, methane emission and populations of ruminal microbes using an in vitro gas production technique. Cablin patchouli herb (CPH), Atractylodes rhizome (AR), Amur Cork-tree (AC) and Cypsum were mixed with the weight ratios of 1:1:1:0.5 and 1:1:1:1 to make up TCMC1 and TCMC2, respectively. Both TCMC were added at level of 25 g/kg of substrate dry matter. In vitro gas production was recorded and methane concentration was determined at 12 and 24 h of incubation. After 24 h, the incubation was terminated and the inoculants were measured for pH, ammonia nitrogen, volatile fatty acids (VFA). Total deoxyribonucleic acid of ruminal microbes was extracted from the inocula, and populations were determined by a real-time quantitative polymerase chain reaction. Populations of total rumen methanogens, protozoa, total fungi, Ruminococcus albus, Fibrobacter succinogenes and Ruminococcus flavefaciens were expressed as a proportion of total rumen bacterial 16S ribosomal deoxyribonucleic acid. Compared with the control, the 2 TCMC decreased (P ≤ 0.05) total VFA concentration, acetate molar proportion, acetate to propionate ratio, gas and methane productions at 12 and 24 h, hydrogen (H) produced and consumed, and methanogens and total fungi populations, while the 2 TCMC increased (P ≤ 0.05) propionate molar proportion. Traditional Chinese medicine compound 1 also decreased (P ≤ 0.05) R. flavefaciens population. From the present study, it is inferred that there is an effect of the TCMC in suppressing methanogenesis, probably mediated via indirect mode by channeling H₂ utilized for methanogenesis to synthesis of propionate and direct action against the rumen microbes involved in methane formation. In addition, the relative methane reduction potential (RMRP) of TCMC2 was superior to that of TCMC1.