反刍与草食动物营养 Ruminant and herbivore nutrition

体外法研究延胡索酸对瘤胃甲烷、氢气产量和挥发性脂肪酸组成的影响

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  • 1. 湖南农业大学动物科学技术学院, 长沙 410128;
    2. 中国科学院亚热带农业生态研究所, 长沙 410125;
    3. 华南农业大学动物科学技术学院, 广州 510642
王荣(1990-),男,河南淮阳人,博士研究生,从事反刍动物营养与饲料科学研究。E-mail:wangrongfd@126.com

收稿日期: 2018-08-17

  网络出版日期: 2019-03-18

基金资助

国家科技计划项目(2018YFD0501800,2016YFD050054);国家自然科学基金项目(31561143009,31472133);湖南省重大专项(2017NK1020);中国科学院青年促进会项目(2016327);中国科学院特聘研究员项目(2018VBA0031)

Effects of Fumarate on Ruminal Methane, Hydrogen Gas Production and Volatile Fatty Acid Composition Using an in Vitro Method

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  • 1. College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, China;
    2. Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China;
    3. College of Animal Science, South China Agricultural University, Guangzhou 510642, China

Received date: 2018-08-17

  Online published: 2019-03-18

摘要

本试验旨在研究不同添加水平的延胡索酸对体外发酵甲烷、氢气产量、挥发性脂肪酸组成以及微生物数量的影响。选用3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。试验采用2×4双因子试验设计,以发酵底物和延胡索酸作为影响因素,其中发酵底物分别是菊苣和玉米粉,延胡索酸设定0、3、6和12 mmol/L 4个添加水平,另外设空白组,即发酵瓶中不添加发酵底物,仅添加0、3、6和12 mmol/L延胡索酸用来矫正数据,每组2个平行,重复测定3次。采用全自动体外模拟瘤胃发酵设备,进行72 h体外模拟瘤胃发酵试验。结果表明:1)玉米粉的72 h产气量、干物质消失率均显著高于菊苣(P<0.05)。延胡索酸显著增加了72 h产气量(P<0.05),但对72 h矫正产气量(去除各个添加水平延胡索酸自身的产气量)、干物质消失率无显著影响(P>0.05)。发酵底物和延胡索酸对72 h产气量和干物质消化率不存在显著交互作用(P>0.05)。2)玉米粉的氢气产量和矫正氢气产量只有在12 h显著高于菊苣(P<0.05),而2种底物在其他时间点的氢气产量和矫正氢气产量无显著差异(P>0.05);玉米粉不同时间点甲烷产量和矫正甲烷产量均显著高于菊苣(P<0.05)。延胡索酸对不同时间点甲烷产量、矫正甲烷产量和氢气产量、氢气矫正产量均无显著影响(P>0.05)。延胡索酸和底物对不同时间点的氢气和甲烷产量不存在显著交互作用(P>0.05)。3)延胡索酸显著增加了乙酸、丙酸和总挥发性脂肪酸浓度(P<0.05),显著降低了乙丙比和pH(P<0.05)。但是,延胡索酸对矫正总挥发性脂肪酸浓度没有显著影响(P>0.05)。4)延胡索酸对微生物数量均没有显著影响(P>0.05)。结果提示,和菊苣相比,玉米粉富含更多易发酵碳水化合物,这有助于产生更多的气体和挥发性脂肪酸。延胡索酸可以增加体外产气量和挥发性脂肪酸浓度主要因为延胡索酸在发酵过程中自身可以代谢产生额外的气体和挥发性脂肪酸,这会使产气量和挥发性脂肪酸浓度增加,而对于发酵底物自身的降解和微生物数量没有显著影响。

本文引用格式

王荣, 文江南, 王敏, 邓近平, 谭支良 . 体外法研究延胡索酸对瘤胃甲烷、氢气产量和挥发性脂肪酸组成的影响[J]. 动物营养学报, 2019 , 31(3) : 1198 -1209 . DOI: 10.3969/j.issn.1006-267x.2019.03.025

Abstract

This study was conducted to investigate the effects of different supplemental levels of fumarate on methane and hydrogen gas production, volatile fatty acids composition and microbial population through in vitro fermentation. Rumen fluid was collected from three adult Xiangdong black goats fitted with permanent rumen cannulation. Using a 2×4 two-factor experimental design, corn flour and chicory were selected as substrates, fumarate had four (0, 3, 6 and 12 mmol/L) final concentrations, and blank groups were set for correcting the results, which were contained no substrate, just contained fumarate at concentrations of 0, 3, 6 or 12 mmol/L, respectively. Each group had 2 replicates, and were replicated 3 times. The 72 h in vitro ruminal incubation was performed using automatic fermentation incubation system. The results showed as follows:1) the 72 h gas production and dry matter degradation of corn flour were significantly higher than those of chicory (P<0.05). Fumarate supplementation had greater 72 h gas production (P<0.05); however, fumarate supplementation did not alter corrected 72 h gas production and dry matter degradation (P>0.05). There is no interaction between substrates and fumarate on 72 h gas production and dry matter degradation (P>0.05). 2) Hydrogen production and corrected hydrogen production of corn flour at 12 h were significantly higher than those of chicory (P<0.05), those at other time points did not have significant differences (P>0.05); methane production and corrected methane production of corn flour at each time points were significantly higher than those of chicory (P<0.05). Fumarate had no significant effect on methane production, corrected methane production, hydrogen production and corrected hydrogen production at each time points (P>0.05). There is no interaction between substrates and fumarate on hydrogen production and methane production (P>0.05). 3) Fumarate supplementation significantly increaed acetate, propionate and total volatile fatty acids concentrations (P<0.05), and significantly decreased pH and acetate to propionate ratio (P<0.05). But fumarate supplementation had no significant effect on corrected total volatile fatty acids concentrations(P>0.05). 4) Fumarate supplementation did not significantly affect microbial population (P>0.05). This study indicates that corn flour contains more fermentable carbohydrates than chicory, which help produce more gas and volatile fatty acids. Fumarate supplementation causes an increase in gas production and total volatile fatty acids concentration, which is mainly due to produce more gas and total volatile fatty acids during fumarate fermentation process, but has no significant effect on substrate self-degradation and microbial population.

参考文献

[1] XU G Q,ZOU W,CHEN X L,et al.Fumaric acid production in Saccharomyces cerevisiae by in silico aided metabolic engineering[J].PLoS One,2012,7(12):e52086.

[2] YIN X,LI J H,SHIN H D,et al.Metabolic engineering in the biotechnological production of organic acids in the tricarboxylic acid cycle of microorganisms:advances and prospects[J].Biotechnology Advances,2015,33(6):830-841.  

[3] KIM S H,MAMUAD L L,KIM D W,et al.Fumarate reductase-producing enterococci reduce methane production in rumen fermentation in vitro[J].Journal of Microbiology and Biotechnology,2016,26(3):558-566.  

[4] ARAÚJO W L,NUNES-NESI A,FERNIE A R.Fumarate:multiple functions of a simple metabolite[J].Phytochemistry,2011,72(9):838-843.  

[5] UNGERFELD E M,KOHN R A,WALLACE R J,et al.A Meta-analysis of fumarate effects on methane production in ruminal batch cultures[J].Journal of Animal Science,2007,85(10):2556-2563.  

[6] UNGERFELD E M.A theoretical comparison between two ruminal electron sinks[J].Frontiers in Microbiology,2013,4:319.

[7] ABRAR A,KONDO M,KITAMURA T,et al.Effect of supplementation of rice bran and fumarate alone or in combination on in vitro rumen fermentation,methanogenesis and methanogens[J].Animal Science Journal,2016,87(3):398-404.  

[8] PISAR?ÍKOVÁ J,VARADYOVÁ Z,MIHALIKOVÁ K,et al.Quantification of organic acids in ruminal in vitro batch culture fermentation supplemented with fumarate using a herb mix as a substrate[J].Canadian Journal of Animal Science,2016,96(1):60-68.  

[9] NEWBOLD C J,LÍPEZ S,NELSON N,et al.Propionate precursors and other metabolic intermediates as possible alternative electron acceptors to methanogenesis in ruminal fermentation in vitro[J].British Journal of Nutrition,2005,94(1):27-35.  

[10] ABDL-RAHMAN M A.In vitro manipulation of rumen fermentation efficiency by fumaric acid-bentonite coupled addition as an alternative to antibiotics[J].Journal of Agricultural Science,2010,2(2):174-180.

[11] PAL K,PATRA A K,SAHOO A,et al.Effect of nitrate and fumarate in Prosopis cineraria and Ailanthus excelsa leaves-based diets on methane production and rumen fermentation[J].Small Ruminant Research,2014,121(2/3):168-174.

[12] KOLVER E S,ASPIN P W,JARVIS G N,et al.Fumarate reduces methane production from pasture fermented in continuous culture[C]//Proceedings of the New Zealand Society of Animal Production.Hamilton:New Zealand Society of Animal Production,2004,155-159.

[13] AOAC.Official methods of analysis of AOAC international[M].18th ed.Washington,D.C.:AOAC International,2005.

[14] MENKE K H,RAAB L,SALEWSKI A,et al.The estimation of the digestibility and metabolizable energy content of ruminant feedingstuffs from the gas production when they are incubated with rumen liquor in vitro[J].The Journal of Agricultural Science,1979,93(1):217-222.  

[15] WANG M,WANG R,YANG S,et al.Effects of three methane mitigation agents on parameters of kinetics of total and hydrogen gas production,ruminal fermentation and hydrogen balance using in vitro technique[J].Animal science Journal,2016,87(2):224-232.  

[16] WANG M,JANSSEN P H,SUN X Z,et al.A mathematical model to describe in vitro kinetics of H2 gas accumulation[J].Animal Feed Science and Technology,2013,184(1/2/3/4):1-16.

[17] WANG M,WANG R,ZHANG X M,et al.Molecular hydrogen generated by elemental magnesium supplementation alters rumen fermentation and microbiota in goats[J].British Journal of Nutrition,2017,118(6):401-410.  

[18] JIAO J Z,W Z,GUAN L L,et al.Postnatal bacterial succession and functional establishment of hindgut in supplemental feeding and grazing goats[J].Journal of Animal Science,2015,93(7):3528-3538.  

[19] DENMAN S E,MCSWEENEY C S.Development of a real-time PCR assay for monitoring anaerobic fungal and cellulolytic bacterial populations within the rumen[J].FEMS Microbiology Ecology,2006,58(3):572-582.  

[20] SYLVESTER J T,KARNATI S K,YU Z T,et al.Development of an assay to quantify rumen ciliate protozoal biomass in cows using real-time PCR[J].The Journal of Nutrition,2004,134(12):3378-3384.  

[21] HOOK S E,NORTHWOOD K S,WRIGHT A D G,et al.Long-term monensin supplementation does not significantly affect the quantity or diversity of methanogens in the rumen of the lactating dairy cow[J].Applied and Environmental Microbiology,2009,75(2):374-380.  

[22] STEVENSON D M,WEIMER P J.Dominance of Prevotella and low abundance of classical ruminal bacterial species in the bovine rumen revealed by relative quantification real-time PCR[J].Applied Microbiology and Biotechnology,2007,75(1):165-174.  

[23] MENKE K H,RAAB L,SALEWSKI A,et al.Estimation of the digestibility and metabolizable energy content of ruminant feedingstuffs from the gas-production when they are incubated with rumen liquor in vitro[J].Journal of Agricultural Science,1979,93:217-222.

[24] SVEINBJORNSSON J,MURPHY M,UDEN P.In vitro evaluation of starch degradation from feeds with or without various heat treatments[J].Animal Feed Science and Technology,2007,132:171-185.

[25] CALLAWAY T R,MARTIN S A.Effects of organic acid and monensin treatment on in vitro mixed ruminal microorganism fermentation of cracked corn[J].Journal of Animal Science,1996,74(8):1982-1989.  

[26] REIS L G,CHAVES A V,WILLIAMS S R O,et al.Comparison of enantiomers of organic acids for their effects on methane production in vitro[J].Animal Production Science,2014,54(9):1345-1349.  

[27] DEMEYER D,HENDERICKX H.Competitive inhibition of "in vitro" methane production by mixed rumen bacteria[J].Archives Internationales de Physiologie et de Biochimie,1967,75(1):157-159.

[28] KITTELMANN S,PINARES-PATINO C S,SEEDORF H,et al.Two different bacterial community types are linked with the low-methane emission trait in sheep[J].Plos One,2014,9:e103171.

[29] JANSSEN P H.Influence of hydrogen on rumen methane formation and fermentation balances through microbial growth kinetics and fermentation thermodynamics[J].Animal Feed Science and Technology,2010,160:1-22.

[30] CASTRO-MONTOYA J,DE CAMPENEERE S,VAN RANST G,et al.Interactions between methane mitigation additives and basal substrates on in vitro methane and VFA production[J].Animal Feed Science and Technology,2012,176(1/2/3/4):47-60.

[31] BAYARU E,SYUHEI K,TOSHIHIKO K.Effect of fumaric acid on methane production,rumen fermentation and digestibility of cattle fed roughage alone[J].Animal Science Journal,2001,72:139-146.

[32] BEAUCHEMIN K A,MCGINN S M.Methane emissions from beef cattle:effects of fumaric acid,essential oil,and canola oil[J].Journal of Animal Science,2006,84(6):1489-1496.  

[33] ENGEL C A R,STRAATHOF A J J,ZIJLMANS T W,et al.Fumaric acid production by fermentation[J].Applied Microbiology and Biotechnology,2008,78(3):379-389.  

[34] REMLING N,RIEDE S,LEBZIEN P,et al.Effects of fumaric acid on rumen fermentation,milk composition and metabolic parameters in lactating cows[J].Journal of Animal Physiology and Animal Nutrition,2014,98(5):968-981.  

[35] MARTIN S A.Manipulation of ruminal fermentation with organic acids:a review[J].Journal of Animal science,1998,76(12):3123-3132.  

[36] WANG M,WANG R,XIE T Y,et al.Shifts in rumen fermentation and microbiota are associated with dissolved ruminal hydrogen concentrations in lactating dairy cows fed fifferent types of carbohydrates[J].The Journal of Nutrition,2016,146(9):1714-1721.  
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