Effect of traditional Chinese medicine compounds on rumen fermentation, methanogenesis and microbial flora in vitro
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摘要
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 16 S 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 H_2 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.
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 16 S 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 H_2 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.
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 16 S 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 H_2 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.
引文
AOAC Official methods of analysis. 18th rev. ed. Gaithersburg, MD, USA:Association of Official Analytic Chemists; 2005.
Benchaar C, Greathead H. Essential oils and opportunities to mitigate enteric methane emissions from ruminants. Anim Feed Sci Technol 2011; 166-167:338-55.
Bodas R, Prieto N, García-González R, Andrés S, Giráldez FJ, López S. Manipulation of rumen fermentation and methane production with plant secondary metabolites. Anim Feed Sci Technol 2012;176:78-93.
Demeyer DI. Quantitative aspects of microbial metabolism in the rumen and hindgut. Rumen microbial metabolism and ruminant digestion. INRA editions.Paris, France:Jouany JP; 1991.
Denman SE, McSweeney CS. Development of a real-time PCR assay for monitoring anaerobic fungal and cellulolytic bacterial populations within the rumen. FEMS Microbiol Ecol 2006;58:572-82.
Denman SE, Tomkins NW, McSweeney CS. Quantitation and diversity analysis of ruminal methanogenic populations in response to the antimethanogenic compound bromochloromethane. FEMS Microbiol Ecol 2007;62:313-22.
Durmic Z, Blache D. Bioactive plants and plant products:effects on animal function,health and welfare. Anim Feed Sci Technol 2012;176:150-62.
Gerber PJ, Hristov AN, Henderson B, Makkar H, Oh J, Lee C, et al. Technical options for the mitigation of direct methane and nitrous oxide emissions from livestock:a review. Animal 2013;7:220-34.
Goel G, Makkar HPS. Methane mitigation from ruminants using tannins and saponins. Trop Anim Health Prod 2012;4:729-39.
Greathead H. Plants and plant extracts for improving animal productivity. Proc Nutr Soc 2003;62:279-90.
Hart KJ, Yanez-Ruiz DR, Duval SM, McEwan NR, Newbold CJ. Plant extracts to manipulate rumen fermentation. Anim Feed Sci Technol 2008;147:8-35.
Hu WL, Liu JX, Ye JA, Wu YM, Guo YQ. Effect of tea saponin on rumen fermentation in vitro. Anim Feed Sci Technol 2005;120:333-9.
Jordan E, Kenny D, Hawkins M, Malone R, Lovett DK, O'Mara FP. Effect of refined soy oil or whole soybeans on intake, methane output, and performance of young bulls. J Anim Sci 2006;84:2418-25.
Kamra DN, Agarwal N, Chaudhary LC. Inhibition of ruminal methanogenesis by tropical plants containing secondary compounds. Int Congr Ser 2006;1293:156-63.
Koike S, Kobayashi Y. Development and use of competitive PCR assays for the rumen cellulolytic bacteria:Fibrobacter succinogenes, Ruminococcus albus and Ruminococcus flavefacies. FEMS Microbiol Lett 2001;204:361-6.
Kumar S, Choudhury PK, Carro MD, Griffith GW, Dagar SS, Puniya M, et al. New aspects and strategies for methane mitigation from ruminants. Appl Microbiol Biotechnol 2014;98:31-44.
Lin B, Lu Y, Wang JH, Liang Q, Liu JX. The effects of combined essential oils along with fumarate on rumen fermentation and methane production in vitro. J Anim Feed Sci 2012;21:198-210.
Liu HW, Tong JM, Zhou DW. Utilization of Chinese herbal feed additives in animal production. Agric Sci China 2011;10:1262-72.
Lu CD. Nutritionally related strategies for organic goat production. Small Rumin Res2011;98:73-82.
Mauricio RM, Mould FL, Dhanoa MS, Owen E, Channa KS, Theodorou MK. A semiautomated in vitro gas production, technique for ruminant feedstuff evaluation.Anim Feed Sci Technol 1999:79:321-30.
McAllister TA, Newbold CJ. Redirecting rumen fermentation to reduce methanogenesis. Aust J Exp Agric 2008;48:7-13.
Menke KH, Raab L, Salewski A, Steingass H, Fritz D, Schneider W. 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 Agric Sci 1979;93:217-22.
Menke KH, Steingass H. Estimation of the energetic feed value obtained from chemical analysis and gas production using rumen fluid. Anim Res Dev1988;28:7-55.
Morgavi D, Martin C, Jouany JP, Ranilla MJ. Rumen protozoa and methanogenesis:not a simple cause-effect relationship. Br J Nutr 2012;107:388-97.
Patra AK, Saxena J. Dietary phytochemicals as rumen modifiers:a review of the effects on microbial populations. Antonie Leeuwenhoek 2009;96:363-75.
Patra AK Enteric methane mitigation technologies for ruminant livestock:a synthesis of current research and future directions. Environ Monit Assess2012:184:1929-52.
Pitt RE, van Kessel JS, Fox DG, Pell AN, Barry MC, van Soest PJ. Prediction of ruminal volatile fatty acids and pH within the net carbohydrate and protein system.J Anim Sci 1996;74:226-44.
SAS, Online doc version 9.1.3. SAS Inst. Inc. Cary, NC, USA:SAS Institute; 2005.
Theodorou MK, Williams BA, Dhanoa MS, McAllan AB, France J. A simple gas production method using a pressure transducer to determine the fermentation kinetics of ruminant feed. Anim Feed Sci Technol 1994;48:185-97.
van Nevel CJ, Demeyer DI. Control of rumen methanogenesis. Environ Monit Assess1996;42:73-97.
van Soest PJ, Robertson JB, Lewis BA. Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. J Dairy Sci1991;74:3583-97.
Wang SP, Wang WJ, Wu M, Zuo FY, Zhou P, Zhang JH, et al. Effect of Chinese medicine prescription on goat:IV. The internal environment parameters of rumen. Chin J Anim Sci 2012b;23:64-7.
Wang SP, Wang WJ, Wu M, Zuo FY, Zhou P, Zhang JH. Effect of Chinese medicine prescription on goat:III. The activity of ruminal celluolytic enzyme and the serum enzyme. Chin J Anim Sci 2012a;21:56-60.
Wang SP, Wang WJ, Zuo FY, Zhou P, Zhang JH. Effect of Chinese medicine prescription on goat:VI. The ruminal degradation characteristics of nutrients from the diet. Chin J Anim Sci 2013;5:47-52.
Wang SP, Wang WJ, Zuo FY, Zhou P, Zhao JJ, Zhang JH. Effect of Chinese medicine prescription on beef cattle in summer:II. Blood gas analysis, concentrations of metabolites and parameters of immune and antioxidant capability in serum.Acta Vet Zootech Sin 2011;42:734-41.
Wang SP, Wang WJ. Effects of dietary supplementation of Chinese herb medicine mixture on rumen fermentation, nutrient digestion and blood profile in goats.S Afr J Anim Sci 2016;46:247-60.
Wang WJ, Wang SP, Zuo FY, Zhou P, Zhang JH. Effect of Chinese Medicine prescription on goat:I. The apparent digestibility of nutrients and the blood routine parameters. Chin J Anim Sci 2012;5:59-62.
Wang WJ, Wang SP, Zuo FY, Zhou P, Zhao JJ, Zhang JH. Effect of Chinese medicine prescription on beef cattle in summer:I. Finishing performance, physiological parameters, serum hormone level and enzymatic activity. Acta Vet Zootech Sin2010;41:1260-7.
Zhang CM, Guo YQ, Yuan ZP, Wu YM, Wang JK, Liu JX, et al. Effect of octadeca carbon fatty acids on microbial fermentation, methanogenesis and microbial flora in vitro. Anim Feed Sci Technol 2008;146:259-69.
Zoetendal EG, Akkermans ADL, De Vos WM. Temperature gradient gel electrophoresis analysis of 16S rRNA from human fecal samples reveals stable and host-specific communities of active bacteria, Appl Environ Microbiol 1998;64.3854-9.
Benchaar C, Greathead H. Essential oils and opportunities to mitigate enteric methane emissions from ruminants. Anim Feed Sci Technol 2011; 166-167:338-55.
Bodas R, Prieto N, García-González R, Andrés S, Giráldez FJ, López S. Manipulation of rumen fermentation and methane production with plant secondary metabolites. Anim Feed Sci Technol 2012;176:78-93.
Demeyer DI. Quantitative aspects of microbial metabolism in the rumen and hindgut. Rumen microbial metabolism and ruminant digestion. INRA editions.Paris, France:Jouany JP; 1991.
Denman SE, McSweeney CS. Development of a real-time PCR assay for monitoring anaerobic fungal and cellulolytic bacterial populations within the rumen. FEMS Microbiol Ecol 2006;58:572-82.
Denman SE, Tomkins NW, McSweeney CS. Quantitation and diversity analysis of ruminal methanogenic populations in response to the antimethanogenic compound bromochloromethane. FEMS Microbiol Ecol 2007;62:313-22.
Durmic Z, Blache D. Bioactive plants and plant products:effects on animal function,health and welfare. Anim Feed Sci Technol 2012;176:150-62.
Gerber PJ, Hristov AN, Henderson B, Makkar H, Oh J, Lee C, et al. Technical options for the mitigation of direct methane and nitrous oxide emissions from livestock:a review. Animal 2013;7:220-34.
Goel G, Makkar HPS. Methane mitigation from ruminants using tannins and saponins. Trop Anim Health Prod 2012;4:729-39.
Greathead H. Plants and plant extracts for improving animal productivity. Proc Nutr Soc 2003;62:279-90.
Hart KJ, Yanez-Ruiz DR, Duval SM, McEwan NR, Newbold CJ. Plant extracts to manipulate rumen fermentation. Anim Feed Sci Technol 2008;147:8-35.
Hu WL, Liu JX, Ye JA, Wu YM, Guo YQ. Effect of tea saponin on rumen fermentation in vitro. Anim Feed Sci Technol 2005;120:333-9.
Jordan E, Kenny D, Hawkins M, Malone R, Lovett DK, O'Mara FP. Effect of refined soy oil or whole soybeans on intake, methane output, and performance of young bulls. J Anim Sci 2006;84:2418-25.
Kamra DN, Agarwal N, Chaudhary LC. Inhibition of ruminal methanogenesis by tropical plants containing secondary compounds. Int Congr Ser 2006;1293:156-63.
Koike S, Kobayashi Y. Development and use of competitive PCR assays for the rumen cellulolytic bacteria:Fibrobacter succinogenes, Ruminococcus albus and Ruminococcus flavefacies. FEMS Microbiol Lett 2001;204:361-6.
Kumar S, Choudhury PK, Carro MD, Griffith GW, Dagar SS, Puniya M, et al. New aspects and strategies for methane mitigation from ruminants. Appl Microbiol Biotechnol 2014;98:31-44.
Lin B, Lu Y, Wang JH, Liang Q, Liu JX. The effects of combined essential oils along with fumarate on rumen fermentation and methane production in vitro. J Anim Feed Sci 2012;21:198-210.
Liu HW, Tong JM, Zhou DW. Utilization of Chinese herbal feed additives in animal production. Agric Sci China 2011;10:1262-72.
Lu CD. Nutritionally related strategies for organic goat production. Small Rumin Res2011;98:73-82.
Mauricio RM, Mould FL, Dhanoa MS, Owen E, Channa KS, Theodorou MK. A semiautomated in vitro gas production, technique for ruminant feedstuff evaluation.Anim Feed Sci Technol 1999:79:321-30.
McAllister TA, Newbold CJ. Redirecting rumen fermentation to reduce methanogenesis. Aust J Exp Agric 2008;48:7-13.
Menke KH, Raab L, Salewski A, Steingass H, Fritz D, Schneider W. 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 Agric Sci 1979;93:217-22.
Menke KH, Steingass H. Estimation of the energetic feed value obtained from chemical analysis and gas production using rumen fluid. Anim Res Dev1988;28:7-55.
Morgavi D, Martin C, Jouany JP, Ranilla MJ. Rumen protozoa and methanogenesis:not a simple cause-effect relationship. Br J Nutr 2012;107:388-97.
Patra AK, Saxena J. Dietary phytochemicals as rumen modifiers:a review of the effects on microbial populations. Antonie Leeuwenhoek 2009;96:363-75.
Patra AK Enteric methane mitigation technologies for ruminant livestock:a synthesis of current research and future directions. Environ Monit Assess2012:184:1929-52.
Pitt RE, van Kessel JS, Fox DG, Pell AN, Barry MC, van Soest PJ. Prediction of ruminal volatile fatty acids and pH within the net carbohydrate and protein system.J Anim Sci 1996;74:226-44.
SAS, Online doc version 9.1.3. SAS Inst. Inc. Cary, NC, USA:SAS Institute; 2005.
Theodorou MK, Williams BA, Dhanoa MS, McAllan AB, France J. A simple gas production method using a pressure transducer to determine the fermentation kinetics of ruminant feed. Anim Feed Sci Technol 1994;48:185-97.
van Nevel CJ, Demeyer DI. Control of rumen methanogenesis. Environ Monit Assess1996;42:73-97.
van Soest PJ, Robertson JB, Lewis BA. Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. J Dairy Sci1991;74:3583-97.
Wang SP, Wang WJ, Wu M, Zuo FY, Zhou P, Zhang JH, et al. Effect of Chinese medicine prescription on goat:IV. The internal environment parameters of rumen. Chin J Anim Sci 2012b;23:64-7.
Wang SP, Wang WJ, Wu M, Zuo FY, Zhou P, Zhang JH. Effect of Chinese medicine prescription on goat:III. The activity of ruminal celluolytic enzyme and the serum enzyme. Chin J Anim Sci 2012a;21:56-60.
Wang SP, Wang WJ, Zuo FY, Zhou P, Zhang JH. Effect of Chinese medicine prescription on goat:VI. The ruminal degradation characteristics of nutrients from the diet. Chin J Anim Sci 2013;5:47-52.
Wang SP, Wang WJ, Zuo FY, Zhou P, Zhao JJ, Zhang JH. Effect of Chinese medicine prescription on beef cattle in summer:II. Blood gas analysis, concentrations of metabolites and parameters of immune and antioxidant capability in serum.Acta Vet Zootech Sin 2011;42:734-41.
Wang SP, Wang WJ. Effects of dietary supplementation of Chinese herb medicine mixture on rumen fermentation, nutrient digestion and blood profile in goats.S Afr J Anim Sci 2016;46:247-60.
Wang WJ, Wang SP, Zuo FY, Zhou P, Zhang JH. Effect of Chinese Medicine prescription on goat:I. The apparent digestibility of nutrients and the blood routine parameters. Chin J Anim Sci 2012;5:59-62.
Wang WJ, Wang SP, Zuo FY, Zhou P, Zhao JJ, Zhang JH. Effect of Chinese medicine prescription on beef cattle in summer:I. Finishing performance, physiological parameters, serum hormone level and enzymatic activity. Acta Vet Zootech Sin2010;41:1260-7.
Zhang CM, Guo YQ, Yuan ZP, Wu YM, Wang JK, Liu JX, et al. Effect of octadeca carbon fatty acids on microbial fermentation, methanogenesis and microbial flora in vitro. Anim Feed Sci Technol 2008;146:259-69.
Zoetendal EG, Akkermans ADL, De Vos WM. Temperature gradient gel electrophoresis analysis of 16S rRNA from human fecal samples reveals stable and host-specific communities of active bacteria, Appl Environ Microbiol 1998;64.3854-9.