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
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摘要
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(P<0.05)。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(P<0.05),且随着山苍子油、茴香油添加浓度的增加呈线性下降趋势(P<0.05);添加400 mg/L山苍子油和茴香油均显著增加了乙酸比例(P<0.05),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(P<0.05);添加200 mg/L茴香油、400 mg/L桉叶油显著提高了丙酸比例(P<0.05),而添加400 mg/L的山苍子油显著降低了丙酸比例(P<0.05),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(P<0.05);添加400 mg/L桉叶油、200 mg/L茴香油显著降低了乙酸/丙酸(P<0.05),添加400 mg/L山苍子油显著增加了乙酸/丙酸(P<0.05),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(P<0.05)。3)与对照组相比,添加400 mg/L茴香油显著降低了NH3-N含量(P<0.05),且随着茴香油添加浓度的增加呈线性下降趋势(P<0.05)。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(P<0.05),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(P<0.05),随着茴香油添加浓度的增加呈线性下降趋势(P<0.05);添加400 mg/L山苍子油、茴香油显著降低了CH4产量(P<0.05),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(P<0.05),随着茴香油添加浓度的增加呈线性下降趋势(P<0.05)。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。
The objective of this experiment was to study the effects of four kinds of plant essential oils(eucalyptus oil,litsea cubeba oil,cinnamon oil and anise oil) on rumen fermentation parameters and methane(CH4) production of mutton sheep in vitro.The ratio of concentrate to forage of the cultivation substrates was60 ∶40,and 0(control),50,100,200 and 400 mg/L of eucalyptus oil,litsea cubeba oil,cinnamon oil or anise oil were added,respectively.There were 3 replicates for each concentration of four plant essential oils.The 24 h gas production,CH4 proportion in gas,pH,volatile fatty acid(VFA) and ammonia nitrogen(NH3-N) concentrations were measured for 24 h simulating rumen fermentation cultivation in vitro.The results showed as follows:1) adding litsea cubeba oil and anise oil significantly affected the rumen fermentation fluid pH in vitro(P<0.05).2) Compared with the control group,adding 400 mg/L litsea cubeba oil and anise oil significantly decreased the total VFA concentration(P<0.05),and showed a linearly decreased trend with the concentrations of litsea cubeba oil and anise oil increased(P<0.05);adding 400 mg/L litsea cubeba oil and anise oil significantly increased the acetic acid proportion(P<0.05),and showed a firstly decreased and then increased quadratically trend with the concentration of litsea cubeba oil and anise oil increased(P < 0.05);adding 200 mg/L anise oil and 400 mg/L eucalyptus oil significantly increased the propionic acid proportion(P< 0.05),adding 400 mg/L litsea cubeba oil significantly decreased the propionic acid proportion(P <0.05),and showed a quadratic curve change trend with the concentrations of eucalyptus oil,litsea cubeba oil and anise oil increased(P<0.05);adding 400 mg/L eucalyptus oil and 200 mg/L anise oil significantly decreased the ratio of acetic acid to propionic acid(P<0.05),adding 400 mg/L litsea cubeba oil significantly increased the ratio of acetic acid to propionic acid(P<0.05),and showed a quadratic curve change trend with the concentrations of eucalyptus oil,anise oil and litsea cubeba oil increased(P<0.05).3) Compared with the control group,adding 400 mg/L litsea cubeba oil and anise oil significantly decreased the NH3-N concentration(P<0.05),and showed a linearly decreased trend with the concentration of anise oil increased(P<0.05).4)Compared with the control group,adding 400 mg/L litsea cubeba oil and anise oil significantly decreased the gas production(P<0.05),and showed a firstly increased and then decreased quadratically trend with the concentration of litsea cubeba oil increased(P<0.05),and showed a linearly decreased trend with the concentration of anise oil increased(P<0.05);adding 400 mg/L litsea cubeba oil and anise oil significantly decreased the CH4 production(P<0.05),and showed a firstly increased and then decreased quadratically trend with the with the concentration increase of litsea cubeba oil increased(P<0.05),and showed a linearly decreased trend with the concentration of anise oil increased.(P<0.05).The results indicate that different plant essential oils have different effects on rumen fermentation parameters and CH4 production in vitro,and which are related to the concentration of essential oils.
The objective of this experiment was to study the effects of four kinds of plant essential oils(eucalyptus oil,litsea cubeba oil,cinnamon oil and anise oil) on rumen fermentation parameters and methane(CH4) production of mutton sheep in vitro.The ratio of concentrate to forage of the cultivation substrates was60 ∶40,and 0(control),50,100,200 and 400 mg/L of eucalyptus oil,litsea cubeba oil,cinnamon oil or anise oil were added,respectively.There were 3 replicates for each concentration of four plant essential oils.The 24 h gas production,CH4 proportion in gas,pH,volatile fatty acid(VFA) and ammonia nitrogen(NH3-N) concentrations were measured for 24 h simulating rumen fermentation cultivation in vitro.The results showed as follows:1) adding litsea cubeba oil and anise oil significantly affected the rumen fermentation fluid pH in vitro(P<0.05).2) Compared with the control group,adding 400 mg/L litsea cubeba oil and anise oil significantly decreased the total VFA concentration(P<0.05),and showed a linearly decreased trend with the concentrations of litsea cubeba oil and anise oil increased(P<0.05);adding 400 mg/L litsea cubeba oil and anise oil significantly increased the acetic acid proportion(P<0.05),and showed a firstly decreased and then increased quadratically trend with the concentration of litsea cubeba oil and anise oil increased(P < 0.05);adding 200 mg/L anise oil and 400 mg/L eucalyptus oil significantly increased the propionic acid proportion(P< 0.05),adding 400 mg/L litsea cubeba oil significantly decreased the propionic acid proportion(P <0.05),and showed a quadratic curve change trend with the concentrations of eucalyptus oil,litsea cubeba oil and anise oil increased(P<0.05);adding 400 mg/L eucalyptus oil and 200 mg/L anise oil significantly decreased the ratio of acetic acid to propionic acid(P<0.05),adding 400 mg/L litsea cubeba oil significantly increased the ratio of acetic acid to propionic acid(P<0.05),and showed a quadratic curve change trend with the concentrations of eucalyptus oil,anise oil and litsea cubeba oil increased(P<0.05).3) Compared with the control group,adding 400 mg/L litsea cubeba oil and anise oil significantly decreased the NH3-N concentration(P<0.05),and showed a linearly decreased trend with the concentration of anise oil increased(P<0.05).4)Compared with the control group,adding 400 mg/L litsea cubeba oil and anise oil significantly decreased the gas production(P<0.05),and showed a firstly increased and then decreased quadratically trend with the concentration of litsea cubeba oil increased(P<0.05),and showed a linearly decreased trend with the concentration of anise oil increased(P<0.05);adding 400 mg/L litsea cubeba oil and anise oil significantly decreased the CH4 production(P<0.05),and showed a firstly increased and then decreased quadratically trend with the with the concentration increase of litsea cubeba oil increased(P<0.05),and showed a linearly decreased trend with the concentration of anise oil increased.(P<0.05).The results indicate that different plant essential oils have different effects on rumen fermentation parameters and CH4 production in vitro,and which are related to the concentration of essential oils.
引文
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[15]张丽英.饲料分析及饲料质量检测技术[M].3版.北京:中国农业大学出版社,2007:140-147.
[16]CAO Y C,YANG H J.Ruminal digestibility and fermentation characteristics in vitro of fenugreek and alfalfa hay combination with or without the inoculation of Neocallimastix sp.YAK11[J].Animal Feed Science and Technology,2011,169(1/2):53-60.
[17]BRODERICK G A,KANG J H.Automated simultaneous determination of ammonia and total amino acids in ruminal fluid and in vitro media[J].Journal of Dairy Science,1980,63(1):64-75.
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[19]CHAVES A V,STANFORD K,DUGAN ME R,et al.Effects of cinnamaldehyde,garlic and juniper berry essential oils on rumen fermentation,blood metabolites,grow th performance,and carcass characteristics of growing lambs[J].Livestock Science 2008,17(2/3):215-224.
[20]BENCHAAR C,MCALLISTER T A,CHOUINARD P Y.Digestion,ruminal fermentation,ciliate protozoal populations,and milk production from dairy cow s fed cinnamaldehyde,quebracho condensed tannin,or Yucca schidigera saponin extracts[J].Journal of Dairy Science,2008,91(12):4765-4777.
[21]YANG W Z,AMETAJ B N,BENCHAAR C,et al.Dose response to cinnamaldehyde supplementation in growing beef heifers:ruminal and intestinal digestion[J].Journal of Animal Science,2010,88(2):680-688.
[22]BUSQUET M,CALSAMIGLIA S,FERRET A,et al.Effects of cinnamaldehyde and garlic oil on rumen microbial fermentation in a dual flow continuous culture[J].Journal of Dairy Science,2005,88(7):2508-2516.
[23]NEWBOLD C J,MCINTOSH F M,WILLIAMS P,et al.Effects of a specific blend of essential oil compounds on rumen fermentation[J].Animal Feed Science and Technology,2004,114(1/2/3/4):105-112.
[24]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.
[25]BUSQUET M,CALSAMIGLIA S,FERRET A,et al.Effects of cinnamaldehyde and garlic oil on rumen microbial fermentation in a dual flow continuous culture[J].Journal of Dairy Science,2005,88(7):2508-2516.
[26]DEVANT M,ANGLADA A,BACH A.Effects of plant extract supplementation on rumen fermentation and metabolism in young Holstein bulls consuming high levels of concentrate[J].Animal Feed Science and Technology,2007,137(1/2):46-57.
[27]王宇星,隋美霞,刘海霞,等.香精油对反刍动物瘤胃发酵的影响[J].饲料博览,2010(2):7-10.
[28]金恩望,卜登攀,王加启,等.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[J].动物营养学报,2013,25(10):2303-2314.
[29]BHATTA R.Reducing enteric methane emission using plant secondary metabolites[M]//SEJIAN V,GAUGHAN J,BAUMGARD L,et al.Climate change impact on livestock:adaptation and mitigation.New Delhi:Springer,2015:273-284.
[30]SZUMACHER-STRABEL M,CIESLAK A.Potential of phytofactors to mitigate rumen ammonia and methane production[J].Journal of Animal and Feed Sciences,2010,19(3):319-337.
[31]OSKOUEIAN E,ABDULLAH N,OSKOUEIAN A.Effects of flavonoids on rumen fermentation activity,methane production,and microbial population[J].BioMed Research International,2013,2013:349129.
[32]COBELLIS G,TRABALZA-MARINUCCI M,YU Z T.Critical evaluation of essential oils as rumen modifiers in ruminant nutrition:a review[J].Science of the Total Environment,2016,545/546:556-568.
[33]PATRA A K,YU Z T.Effects of essential oils on methane production and fermentation by,and abundance and diversity of,rumen microbial populations[J].Applied and Environmental Microbiology,2012,78(12):4271-4280.
[34]JAYANEGARA A,GOEL G,MAKKAR H P S,et al.Divergence between purified hydrolysable and condensed tannin effects on methane emission,rumen fermentation and microbial population in vitro[J].Animal Feed Science and Technology,2015,209:60-68.
[35]PINSKI B,GNAL M,ABUGHAZALEH A A.The effects of essential oil and condensed tannin on fermentation and methane production under in vitro conditions[J].Animal Production Science,2015,56(10):1707-1713.
[36]林波,纪苗苗,梁权,等.肉桂油和牛至油及其主要成分对体外瘤胃发酵和甲烷产生的影响[J].中国兽医学报,2011,31(2):279-282,287.
[37]PATRA A K,SAXENA J.A new perspective on the use of plant secondary metabolites to inhibit methanogenesis in the Rumen[J].Phytochemistry,2010,41(11/12):1198-1222.
[2]KIRSCHKE S,BOUSQUET P,CIAIS P,et al.Three decades of global methane sources and sinks[J].Nature Geoscience,2013,6(10):813-823.
[3]LASSEY K R.Livestock methane emission and its perspective in the global methane cycle[J].Australian Journal of Experimental Agriculture,2008,48(2):114-118.
[4]ECKARD R J,GRAINGER C,DE KLEIN C A M.Options for the abatement of methane and nitrous oxide from ruminant production:a review[J].Livestock Science,2010,130(1/2/3):47-56.
[5]SALLAMS,BUENO I C S,BRIGIDE P,et al.Investigation of potential newopportunities for plant extracts on rumen microbial fermentation in vitro[J].Options Méditerranéennes Série A Séminai,2009,85:255-260.
[6]MOLERO R,IBARS M,CALSAMIGLIA S,et al.Effects of a specific blend of essential oil compounds on dry matter and crude protein degradability in heifers fed diets with different forage to concentrate ratios[J].Animal Feed Science and Technology,2004,114(1/2/3/4):91-104.
[7]CARDOZO P W,CALSAMIGLIA S,FERRET A,et al.Screening for the effects of natural plant extracts at different pH on in vitro rumen microbial fermentation of a high-concentrate diet for beef cattle[J].Journal of Animal Science,2005,83(11):2572-2579.
[8]BUSQUET M,CALSAMIGLIA S,FERRET A,et al.Plant extracts affect in vitro rumen microbial fermentation[J].Journal of Dairy Science,2006,89(2):761-771.
[9]CASTILLEJOS L,CALSAMIGLIA S,FERRET A.Effect of essential oil active compounds on rumen microbial fermentation and nutrient flowin in vitro systems[J].Journal of Dairy Science,2006,89(7):2649-2658.
[10]CALSAMIGLIA S,BUSQUET M,CARDOZO P W,et al.invited review:essential oils as modifiers of rumen microbial fermentation[J].Journal of Dairy Science,2007,90(6):2580-2595.
[11]LI Y L,HE ML,LI C,et al.Effects of wheat dried distillers grains with solubles and cinnamaldehyde on in vitro fermentation and protein degradation using the Rusitec technique[J].Archives of Animal Nutrition,2012,66(2):131-148.
[12]MENKE K H,RAAB L,SALEWSKI A,et al.The estimation of the digestibility and metabolizable energy content of ruminant feeding stuffs from the gas production when they are incubated with rumen liquor in vitro[J].The Journal of Agricultural Science,1979,93(1):217-222.
[13]VAN SOEST P J,ROBERTSON J B,LEWIS B A.Methods for dietary fiber,neutral detergent fiber,and nonstarch polysaccharides in relation to animal nutrition[J].Journal of Dairy Science,1991,74(10):3583-3597.
[14]AOAC.Official methods of analysis[S].16th ed.Arlington,VA:Association of Official Analytical Chemists,1995.
[15]张丽英.饲料分析及饲料质量检测技术[M].3版.北京:中国农业大学出版社,2007:140-147.
[16]CAO Y C,YANG H J.Ruminal digestibility and fermentation characteristics in vitro of fenugreek and alfalfa hay combination with or without the inoculation of Neocallimastix sp.YAK11[J].Animal Feed Science and Technology,2011,169(1/2):53-60.
[17]BRODERICK G A,KANG J H.Automated simultaneous determination of ammonia and total amino acids in ruminal fluid and in vitro media[J].Journal of Dairy Science,1980,63(1):64-75.
[18]KHATERI N,AZIZI O,JAHANI-AZIZABADI H.Effects of a specific blend of essential oils on apparent nutrient digestion,rumen fermentation and rumen microbial populations in sheep fed a 50∶50 alfalfa hay:concentrate diet[J].Asian-Australasian Journal of Animal Sciences,2017,30(3):370-378.
[19]CHAVES A V,STANFORD K,DUGAN ME R,et al.Effects of cinnamaldehyde,garlic and juniper berry essential oils on rumen fermentation,blood metabolites,grow th performance,and carcass characteristics of growing lambs[J].Livestock Science 2008,17(2/3):215-224.
[20]BENCHAAR C,MCALLISTER T A,CHOUINARD P Y.Digestion,ruminal fermentation,ciliate protozoal populations,and milk production from dairy cow s fed cinnamaldehyde,quebracho condensed tannin,or Yucca schidigera saponin extracts[J].Journal of Dairy Science,2008,91(12):4765-4777.
[21]YANG W Z,AMETAJ B N,BENCHAAR C,et al.Dose response to cinnamaldehyde supplementation in growing beef heifers:ruminal and intestinal digestion[J].Journal of Animal Science,2010,88(2):680-688.
[22]BUSQUET M,CALSAMIGLIA S,FERRET A,et al.Effects of cinnamaldehyde and garlic oil on rumen microbial fermentation in a dual flow continuous culture[J].Journal of Dairy Science,2005,88(7):2508-2516.
[23]NEWBOLD C J,MCINTOSH F M,WILLIAMS P,et al.Effects of a specific blend of essential oil compounds on rumen fermentation[J].Animal Feed Science and Technology,2004,114(1/2/3/4):105-112.
[24]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.
[25]BUSQUET M,CALSAMIGLIA S,FERRET A,et al.Effects of cinnamaldehyde and garlic oil on rumen microbial fermentation in a dual flow continuous culture[J].Journal of Dairy Science,2005,88(7):2508-2516.
[26]DEVANT M,ANGLADA A,BACH A.Effects of plant extract supplementation on rumen fermentation and metabolism in young Holstein bulls consuming high levels of concentrate[J].Animal Feed Science and Technology,2007,137(1/2):46-57.
[27]王宇星,隋美霞,刘海霞,等.香精油对反刍动物瘤胃发酵的影响[J].饲料博览,2010(2):7-10.
[28]金恩望,卜登攀,王加启,等.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[J].动物营养学报,2013,25(10):2303-2314.
[29]BHATTA R.Reducing enteric methane emission using plant secondary metabolites[M]//SEJIAN V,GAUGHAN J,BAUMGARD L,et al.Climate change impact on livestock:adaptation and mitigation.New Delhi:Springer,2015:273-284.
[30]SZUMACHER-STRABEL M,CIESLAK A.Potential of phytofactors to mitigate rumen ammonia and methane production[J].Journal of Animal and Feed Sciences,2010,19(3):319-337.
[31]OSKOUEIAN E,ABDULLAH N,OSKOUEIAN A.Effects of flavonoids on rumen fermentation activity,methane production,and microbial population[J].BioMed Research International,2013,2013:349129.
[32]COBELLIS G,TRABALZA-MARINUCCI M,YU Z T.Critical evaluation of essential oils as rumen modifiers in ruminant nutrition:a review[J].Science of the Total Environment,2016,545/546:556-568.
[33]PATRA A K,YU Z T.Effects of essential oils on methane production and fermentation by,and abundance and diversity of,rumen microbial populations[J].Applied and Environmental Microbiology,2012,78(12):4271-4280.
[34]JAYANEGARA A,GOEL G,MAKKAR H P S,et al.Divergence between purified hydrolysable and condensed tannin effects on methane emission,rumen fermentation and microbial population in vitro[J].Animal Feed Science and Technology,2015,209:60-68.
[35]PINSKI B,GNAL M,ABUGHAZALEH A A.The effects of essential oil and condensed tannin on fermentation and methane production under in vitro conditions[J].Animal Production Science,2015,56(10):1707-1713.
[36]林波,纪苗苗,梁权,等.肉桂油和牛至油及其主要成分对体外瘤胃发酵和甲烷产生的影响[J].中国兽医学报,2011,31(2):279-282,287.
[37]PATRA A K,SAXENA J.A new perspective on the use of plant secondary metabolites to inhibit methanogenesis in the Rumen[J].Phytochemistry,2010,41(11/12):1198-1222.