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RESEARCH ARTICLE

Effects of the orange lemma (rob1) mutant line of barley cv. ‘Optic’ compared with its wild-type on the ruminal microbiome and fermentation tested with the rumen simulation technique

F. Klevenhusen https://orcid.org/0000-0001-6015-5459 A C , C. Emsenhuber B , H. Grausgruber https://orcid.org/0000-0003-4790-0922 B , R. M. Petri A and Q. Zebeli A C
+ Author Affiliations
- Author Affiliations

A Institute of Animal Nutrition and Functional Plant Compounds, Department for Farm Animals and Veterinary Public Health, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210 Vienna, Austria.

B Department of Crop Sciences, University of Natural Resources and Life Sciences, Vienna, Konrad-Lorenz-Str. 24, 3430 Tulln, Austria.

C Corresponding authors. Email: fenjaklevenhusen@aol.com; qendrim.zebeli@vetmeduni.ac.at

Crop and Pasture Science 70(9) 789-799 https://doi.org/10.1071/CP18288
Submitted: 20 June 2018  Accepted: 22 July 2019   Published: 27 September 2019

Abstract

The use of cereals as forage crops is limited due to the high lignin content in the cell walls reducing nutrient digestibility. Recent research has focused on reducing lignification in forage crops through gene mutations. This study investigated the ruminal fermentation characteristics of a barley mutation (orange lemma), which is associated with a lower lignin content, using the in vitro ruminal fermentation system (RUSITEC). Two-rowed spring barley cv. ‘Optic’ and its ethyl methane sulfonate (EMS)-induced orange lemma (rob1) mutant line were harvested at both stem elongation and early fruit development and incubated in the RUSITEC system. Gas production, concentrations of short-chain fatty acids (SCFA) and ammonia and the nutrient degradation of the plants after 48 h incubation were investigated. Additional samples were analysed for microbial composition using MiSeq sequencing technology. In general, acid detergent lignin (ADL) was higher at early grain filling than stem elongation. ADL was lower in the mutant line than in the wild type at both stem elongation (13.9% vs 18.5%) and early grain development (26.0% vs 28.6%; dry matter basis). This was reflected in increased ruminal degradation of neutral detergent fibre (61.7% vs 53.7%; P < 0.001) when harvested at stem elongation, but not at the later stage. In contrast, methane formation was lower with rob1 than ‘Optic’ (P = 0.002), especially when harvested at stem elongation. No difference was seen in protein degradation between the barley genotypes. The fermentation SCFA profile did not differ between barley genotypes when harvested at stem elongation, but at early fruit development more acetate and less butyrate was produced with rob1. Microbial species richness was lower when barley was incubated at stem elongation compared to fruit development (P < 0.001), which was especially pronounced with rob1 (P = 0.026). The abundance of Bacteroidetes, Synergistetes and Tenericutes was lower when plants harvested at early grain development were incubated compared to the stem elongation stage, whereas the abundance of Cyanobacteria, Elusimicrobia, Fusobacteria, Lentisphaerae, Proteobacteria, Verrucomicrobia and WPS-2 was higher (P < 0.001). In conclusion, most fermentation parameters were affected by vegetation stage and related changes in nutrient composition. However, additional effects of barley genotype were seen on the rumen microbial community structure, SCFA profile and methane production.

Additional keywords: 16S rRNA gene, incubation, methane, rumen microbiota, short-chain fatty acids.


References

Andries JI, Buysse FX, De Brabander DL, Cottyn BG (1987) Isoacids in ruminant nutrition: their role in ruminal and intermediary metabolism and possible influences on performances – a review. Animal Feed Science and Technology 18, 169–180.
Isoacids in ruminant nutrition: their role in ruminal and intermediary metabolism and possible influences on performances – a review.Crossref | GoogleScholarGoogle Scholar |

Avguštin G, Wallace RJ, Flint HJ (1997) Phenotypic diversity among ruminal isolates of Prevotella ruminicola: proposal of Prevotella brevis sp. nov., Prevotella byantii sp. nov., and Prevotella albensis sp. nov. and redefinition of Prevotella ruminicola. International Journal of Systematic and Evolutionary Microbiology 47, 284–288.

Baron VS, Dick AC, McCartney D, Basarab JA, Okine EK (2006) Carrying capacity, utilization, and weathering of swathed whole plant barley. Agronomy Journal 98, 714–721.
Carrying capacity, utilization, and weathering of swathed whole plant barley.Crossref | GoogleScholarGoogle Scholar |

Belanche A, Kingston-Smith AH, Griffith GW, Newbold CJ (2019) A multi-kingdom study reveals the plasticity of the rumen microbiota in response to a shift from non-grazing to grazing diets in sheep. Frontiers in Microbiology 10, 122
A multi-kingdom study reveals the plasticity of the rumen microbiota in response to a shift from non-grazing to grazing diets in sheep.Crossref | GoogleScholarGoogle Scholar | 30853943PubMed |

Bell LW, Moore AD, Kirkegaard JA (2014) Evolution in crop–livestock integration systems that improve farm productivity and environmental performance in Australia. European Journal of Agronomy 57, 10–20.
Evolution in crop–livestock integration systems that improve farm productivity and environmental performance in Australia.Crossref | GoogleScholarGoogle Scholar |

Bergman EN (1990) Energy contribution of VFA from the gastrointestinal tract in various species. APS Physiological Reviews 70, 567–590.
Energy contribution of VFA from the gastrointestinal tract in various species.Crossref | GoogleScholarGoogle Scholar |

Bokulich NA, Subramanian S, Faith JJ, Gevers D, Gordon JI, Knight R, Mills DA, Caporaso JG (2013) Quality-filtering vastly improves diversity estimates from Illumina amplicon sequencing. Nature Methods 10, 57–59.
Quality-filtering vastly improves diversity estimates from Illumina amplicon sequencing.Crossref | GoogleScholarGoogle Scholar | 23202435PubMed |

Caldwell DG, McCallum N, Shaw P, Muehlbauer GJ, Marshall DF, Waugh R (2004) A structured mutant population for forward and reverse genetics in barley (Hordeum vulgare L.). The Plant Journal 40, 143–150.
A structured mutant population for forward and reverse genetics in barley (Hordeum vulgare L.).Crossref | GoogleScholarGoogle Scholar | 15361148PubMed |

Caporaso JG, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK, Fierer N, Peña AG, Goodrich JK, Gordon JI, Huttley GA, Kelley ST, Knights D, Koenig JE, Ley RE, Lozupone CA, McDonald D, Muegge BD, Pirrung M, Reeder J, Sevinsky JR, Turnbaugh PJ, Walters WA, Widmann J, Yatsunenko T, Zaneveld J, Knight R (2010) QIIME allows analysis of high-throughput community sequencing data. Nature Methods 7, 335–336.
QIIME allows analysis of high-throughput community sequencing data.Crossref | GoogleScholarGoogle Scholar | 20383131PubMed |

Chapple C, Ladisch M, Meilan R (2007) Loosening lignin’s grip on biofuel production. Nature Biotechnology 25, 746–748.
Loosening lignin’s grip on biofuel production.Crossref | GoogleScholarGoogle Scholar | 17621299PubMed |

Chen F, Dixon RA (2007) Lignin modification improves fermentable sugar yields for biofuel production. Nature Biotechnology 25, 759–761.
Lignin modification improves fermentable sugar yields for biofuel production.Crossref | GoogleScholarGoogle Scholar | 17572667PubMed |

Cheng Y, Wang Y, Li Y, Zhang Y, Liu T, Wang Y, Sharpton TJ, Zhu W (2017) Progressive colonization of bacteria and degradation of rice straw in the rumen by Illumina sequencing. Frontiers in Microbiology 8, 2165
Progressive colonization of bacteria and degradation of rice straw in the rumen by Illumina sequencing.Crossref | GoogleScholarGoogle Scholar | 29163444PubMed |

Collar C, Wright S, Robinson P, Putnam D (2004) Effect of harvest timing on yield and quality of small grain forage. In ‘Proceedings of the 34th California Alfalfa and 2004 National Alfalfa Symposium’. 13–15 December 2004, San Diego, CA, USA. pp. 141–148. (University of California: Davis, CA, USA)

Comtet-Marre S, Parisot N, Lepercq P, Chaucheyras-Durand F, Mosoni P, Peyretaillade E, Bayat AR, Shingfield KJ, Peyret P, Forano E (2017) Metatranscriptomics reveals the active bacterial and eukaryotic fibrolytic communities in the rumen of dairy cow fed a mixed diet. Frontiers in Microbiology 8, 67
Metatranscriptomics reveals the active bacterial and eukaryotic fibrolytic communities in the rumen of dairy cow fed a mixed diet.Crossref | GoogleScholarGoogle Scholar | 28197133PubMed |

Cotta MA, Hespel RB (1986) Proteolytic activity of the ruminal bacterium Butyrivibrio fibrisolvens. Applied and Environmental Microbiology 52, 51–58.

Dai X, Tian Y, Li J, Su X, Wang X, Zhao S, Liu L, Luo Y, Liu D, Zheng H, Wang J, Dong Z, Hu S, Huang L (2015) Metatranscriptomic analyses of plant cell wall polysaccharide degradation by microorganisms in the cow rumen. Applied and Environmental Microbiology 81, 1375–1386.
Metatranscriptomic analyses of plant cell wall polysaccharide degradation by microorganisms in the cow rumen.Crossref | GoogleScholarGoogle Scholar | 25501482PubMed |

de Menezes AB, Lewis E, O’Donovan M, O’Neill BF, Clipson N, Doyle EM (2011) Microbiome analysis of dairy cows fed pasture or total mixed ration diets. FEMS Microbiology Ecology 78, 256–265.
Microbiome analysis of dairy cows fed pasture or total mixed ration diets.Crossref | GoogleScholarGoogle Scholar | 21671962PubMed |

Edgar RC (2010) Search and clustering orders of magnitude faster than BLAST. Bioinformatics 26, 2460–2461.
Search and clustering orders of magnitude faster than BLAST.Crossref | GoogleScholarGoogle Scholar | 20709691PubMed |

Entz MH, Baron VS, Carr PM, Meyer DW, Smith SR, McCaughey WP (2002) Potential of forages to diversify cropping systems in the Northern Great Plains. Agronomy Journal 94, 240–250.
Potential of forages to diversify cropping systems in the Northern Great Plains.Crossref | GoogleScholarGoogle Scholar |

Eticha F, Berghofer E, Grausgruber H (2009) Utilization of barley (Hordeum vulgare L.) landraces in the highlands of West Shewa, Ethiopia. Plant Genetic Resources 7, 154–162.
Utilization of barley (Hordeum vulgare L.) landraces in the highlands of West Shewa, Ethiopia.Crossref | GoogleScholarGoogle Scholar |

Fisher LJ, Lessard JR, Lodge GA (1972) Whole crop barley as conserved forage for lactating cows. Canadian Journal of Animal Science 52, 497–504.
Whole crop barley as conserved forage for lactating cows.Crossref | GoogleScholarGoogle Scholar |

Hadjichristodoulou A (1979) Sanokrithi 74 – a hooded barley for hay. Technical Bulletin 28, Agricultural Research Institute, Nicosia, Cyprus.

Harlan HV (1920) Smooth-awned barleys. Journal - American Society of Agronomy 12, 205–208.
Smooth-awned barleys.Crossref | GoogleScholarGoogle Scholar |

Hatew B, Cone JW, Pellikaan WF, Podesta SC, Bannink A, Hendriks WH, Dijkstra J (2015) Relationship between in vitro and in vivo methane production measured simultaneously with different dietary starch sources and starch levels in dairy cattle. Animal Feed Science and Technology 202, 20–31.
Relationship between in vitro and in vivo methane production measured simultaneously with different dietary starch sources and starch levels in dairy cattle.Crossref | GoogleScholarGoogle Scholar |

Henderson G, Cox F, Ganesh S, Jonker A, Young W, Janssen PH (2015) Rumen microbial community composition varies with diet and host, but a core microbiome is found across a wide geographical range. Scientific Reports 5, 14567
Rumen microbial community composition varies with diet and host, but a core microbiome is found across a wide geographical range.Crossref | GoogleScholarGoogle Scholar | 26449758PubMed |

Jin W, Wang Y, Li Y, Cheng Y, Zhu W (2018) Temporal changes of the bacterial community colonizing wheat straw in the cow rumen. Anaerobe 50, 1–8.
Temporal changes of the bacterial community colonizing wheat straw in the cow rumen.Crossref | GoogleScholarGoogle Scholar | 29330119PubMed |

Jung H-JG, Samac DA, Sarath G (2012) Review: modifying crops to increase cell wall digestibility. Plant Science 185–186, 65–77.
Review: modifying crops to increase cell wall digestibility.Crossref | GoogleScholarGoogle Scholar |

Kasperowicz A, Stan-Glasek K, Guczynska W, Piknova M, Pristas P, Nigutova K, Javorsky P, Michalowski T (2009) Sucrose phosphorylase of the rumen bacterium Pseudobutyrivibrio ruminis strain A. Journal of Applied Microbiology 107, 812–820.
Sucrose phosphorylase of the rumen bacterium Pseudobutyrivibrio ruminis strain A.Crossref | GoogleScholarGoogle Scholar | 19320946PubMed |

Khorasani GR, Jedel PE, Helm JH, Kennelly JJ (1997) Influence of stage of maturity on yield components and chemical composition of cereal grain silages. Canadian Journal of Animal Science 77, 259–267.
Influence of stage of maturity on yield components and chemical composition of cereal grain silages.Crossref | GoogleScholarGoogle Scholar |

Klevenhusen F, Petri RM, Kleefisch M-T, Khiaosa-ard R, Metzler-Zebeli BU, Zebeli Q (2017) Changes in fibre-adherent and fluid-associated microbial communities and fermentation profiles in the rumen of cattle fed diets differing in hay quality and concentrate amount. FEMS Microbioly Ecology 93,
Changes in fibre-adherent and fluid-associated microbial communities and fermentation profiles in the rumen of cattle fed diets differing in hay quality and concentrate amount.Crossref | GoogleScholarGoogle Scholar |

Kong Y, Teather R, Forster R (2010) Composition, spatial distribution, and diversity of the bacterial communities in the rumen of cows fed different forages. FEMS Microbiology Ecology 74, 612–622.
Composition, spatial distribution, and diversity of the bacterial communities in the rumen of cows fed different forages.Crossref | GoogleScholarGoogle Scholar | 21044097PubMed |

Lundqvist U, Franckowiak JD (2003) Diversity of barley mutants. In ‘Diversity in barley (Hordeum vulgare)’. (Eds R von Bothmer, T van Hintum, H Knüpffer, K Sato) pp. 77–96. (Elsevier: Amsterdam)

Mannerkorpi P, Taube F (1995) Feeding value of barley plants as related to stage of maturity 1. Morphological and chemical composition and in vitro digestibility. Acta Agriculturæ Scandinavica. Section A, Animal Science 45, 147–152.
Feeding value of barley plants as related to stage of maturity 1. Morphological and chemical composition and in vitro digestibility.Crossref | GoogleScholarGoogle Scholar |

McCartney D, Fraser J, Ohama A (2008) Annual cool season crops for grazing by beef cattle. A Canadian review. Canadian Journal of Animal Science 88, 517–533.
Annual cool season crops for grazing by beef cattle. A Canadian review.Crossref | GoogleScholarGoogle Scholar |

Münnich M, Khol-Parisini A, Klevenhusen F, Metzler-Zebeli BU, Zebeli Q (2018) Graded replacement of maize grain with molassed sugar beet pulp modulated ruminal microbial community and fermentation profile in vitro. Journal of the Science of Food and Agriculture 98, 991–997.
Graded replacement of maize grain with molassed sugar beet pulp modulated ruminal microbial community and fermentation profile in vitro.Crossref | GoogleScholarGoogle Scholar | 28722751PubMed |

Nocek JE, Russell JB (1988) Protein and energy as an integrated system. Relationship of ruminal protein and carbohydrate availability to microbial synthesis and milk production. Journal of Dairy Science 71, 2070–2107.
Protein and energy as an integrated system. Relationship of ruminal protein and carbohydrate availability to microbial synthesis and milk production.Crossref | GoogleScholarGoogle Scholar |

Owings D, Franckowiak J, Nudell R (2006) Forage quality of barley hay. In ‘ASA-CSSA-SSSA 2006 International Meetings’. 12–16 November 2006, Indianapolis, IN, USA. Abstract 266-15. (American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America: Madison, WI, USA)

Peterson J, Garges S, Giovanni M, McInnes P, Wang L, Schloss JA, Bonazzi V, McEwen JE, Wetterstrand KA, Deal C, Baker CC, Di Francesco V, Howcroft TK, Karp RW, Lunsford RD, Wellington CR, Belachew T, Wright M, Giblin C, David H, Mills M, Salomon R, Mullins C, Akolkar B, Begg L, Davis C, Grandison L, Humble M, Khalsa J, Little AR, Peavy H, Pontzer C, Portnoy M, Sayre MH, Starke-Reed P, Zakhari S, Read J, Watson B, Guyer M (2009) The NIH human microbiome project. Genome Research 19, 2317–2323.
The NIH human microbiome project.Crossref | GoogleScholarGoogle Scholar | 19819907PubMed |

Petri RM, Schwaiger T, Penner GB, Beauchemin KA, Forster RJ, McKinnon JJ, McAllister TA (2013) Changes in the rumen epimural bacterial diversity of beef cattle as affected by diet and induced ruminal acidosis. Applied and Environmental Microbiology 79, 3744–3755.
Changes in the rumen epimural bacterial diversity of beef cattle as affected by diet and induced ruminal acidosis.Crossref | GoogleScholarGoogle Scholar | 23584771PubMed |

Pitta DW, Indugu N, Baker L, Vecchiarelli B, Attwood G (2018) Symposium review: understanding diet–microbe interactions to enhance productivity of dairy cows. Journal of Dairy Science 101, 7661–7679.
Symposium review: understanding diet–microbe interactions to enhance productivity of dairy cows.Crossref | GoogleScholarGoogle Scholar | 29859694PubMed |

Poehlman JM (1985) Adaptation and distribution. In ‘Barley’. (Ed. DC Rasmusson) pp. 1–17. (American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America: Madison, WI, USA)

Quast C, Pruesse E, Yilmaz P, Gerken J, Schweer T, Yarza P, Peplies J, Glöckner FO (2012) The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Research 41, D590–D596.
The SILVA ribosomal RNA gene database project: improved data processing and web-based tools.Crossref | GoogleScholarGoogle Scholar | 23193283PubMed |

Qumar M, Khiaosa-ard R, Pourazad P, Wetzels SU, Klevenhusen F, Kandler W, Aschenbach JR, Zebeli Q (2016) Evidence of in vivo absorption of lactate and modulation of short chain fatty acid absorption from the reticulorumen of non-lactating cattle fed high concentrate diets. PLoS One 11, e0164192
Evidence of in vivo absorption of lactate and modulation of short chain fatty acid absorption from the reticulorumen of non-lactating cattle fed high concentrate diets.Crossref | GoogleScholarGoogle Scholar | 27716806PubMed |

Radchuk VV, Borisjuk L, Sreenivasulu N, Merx K, Mock HP, Rolletschek H, Wobus U, Weschke W (2009) Spatiotemporal profiling of starch biosynthesis and degradation in the developing barley grain. Plant Physiology 150, 190–204.
Spatiotemporal profiling of starch biosynthesis and degradation in the developing barley grain.Crossref | GoogleScholarGoogle Scholar | 19321714PubMed |

Sattler SE, Funnell-Harris DL, Pedersen JF (2010) Brown midrib mutations and their importance to the utilization of maize, sorghum, and pearl millet lignocellulosic tissues. Plant Science 178, 229–238.
Brown midrib mutations and their importance to the utilization of maize, sorghum, and pearl millet lignocellulosic tissues.Crossref | GoogleScholarGoogle Scholar |

Shen H, Fu C, Xiao X, Ray T, Tang Y, Wang Z, Chen F (2009) Developmental control of lignification in stems of lowland switchgrass variety Alamo and the effects on saccharification efficiency. BioEnergy Research 2, 233–245.
Developmental control of lignification in stems of lowland switchgrass variety Alamo and the effects on saccharification efficiency.Crossref | GoogleScholarGoogle Scholar |

Shinkai T, Kobayashi Y (2007) Localization of ruminal cellulolytic bacteria on plant fibrous materials as determined by fluorescence in situ hybridization and real-time PCR. Applied and Environmental Microbiology 73, 1646–1652.
Localization of ruminal cellulolytic bacteria on plant fibrous materials as determined by fluorescence in situ hybridization and real-time PCR.Crossref | GoogleScholarGoogle Scholar | 17209077PubMed |

Soliva CR, Hess HD (2007) Measuring methane emission of ruminants by in vitro and in vivo techniques. In ‘Measuring methane production from ruminants’. (Eds HPS Makkar, PE Vercoe) pp. 15–31. (Springer: Dordrecht, The Netherlands)

Tashi N, Yawei T, Xingquan Z (2013) Food preparation from hulless barley in Tibet. In ‘Advance in barley sciences. Proceedings of the 11th International Barley Genetics Symposium’. (Eds G Zhang, C Li, X Liu) pp. 151–158. (Springer: Dordrecht, The Netherlands) 10.1007/978-94-007-4682-4_13

Todd AL, Bowman JGP, Surber LMM, Thompson MA, Kincheloe JJ, McDonnell MF, Hensleigh PF (2003) Effect of barley varieties harvested for forage on backgrounding steer performance and diet digestibility. Proceedings, Western Section. American Society of Animal Science 54, 389–392.

Van Gylswyk NO, Hippe H, Rainey FA (1996) Pseudobutyrivibrio ruminis gen. nov., sp. nov., a butyrate-producing bacterium from the rumen that closely resembles Butyrivibrio fibrisolvens in phenotype. Internation Journal of Systemic and Evolutionary Microbiology 46, 559–563.

VDLUFA (Verband Deutscher Landwirtschaftlicher Untersuchungs- und Forschungsanstalten) (2012) ‘Handbuch der Landwirtschaftlichen Versuchs- und Untersuchungsmethodik (VDLUFA-Methodenbuch), Bd. III. Die chemische Untersuchung von Futtermitteln.’ (VDLUFA: Darmstadt)

Vogel KP, Jung H-JG (2001) Genetic modification of herbaceous plants for feed and fuel. Critical Reviews in Plant Sciences 20, 15–49.
Genetic modification of herbaceous plants for feed and fuel.Crossref | GoogleScholarGoogle Scholar |

Wallsten J, Bertilsson J, Nadeau E, Martinsson K (2010) Digestibility of whole-crop barley and oat silages in dairy heifers. Animal 4, 432–438.
Digestibility of whole-crop barley and oat silages in dairy heifers.Crossref | GoogleScholarGoogle Scholar | 22443947PubMed |

Weatherburn M (1967) Phenol–hypochlorite reaction for determination of ammonia. Analytical Chemistry 39, 971–974.
Phenol–hypochlorite reaction for determination of ammonia.Crossref | GoogleScholarGoogle Scholar |

Weschke W, Panitz R, Sauer N, Wang Q, Neubohn B, Weber H, Wobus U (2000) Sucrose transport into barley seeds: molecular characterization of two transporters and implications for seed development and starch accumulation. The Plant Journal 21, 455–467.
Sucrose transport into barley seeds: molecular characterization of two transporters and implications for seed development and starch accumulation.Crossref | GoogleScholarGoogle Scholar | 10758497PubMed |

Wetzels SU, Eger M, Burmester M, Kreienbrock L, Abdulmawjood A, Pinior B, Wagner M, Breves G, Mann E (2018) The application of rumen simulation technique (RUSITEC) for studying dynamics of the bacterial community and metabolome in rumen fluid and the effects of a challenge with Clostridium perfringens. PLoS One 13, e0192256
The application of rumen simulation technique (RUSITEC) for studying dynamics of the bacterial community and metabolome in rumen fluid and the effects of a challenge with Clostridium perfringens.Crossref | GoogleScholarGoogle Scholar | 29415046PubMed |

Yilmaz P, Parfrey LW, Yarza P, Gerken J, Pruesse E, Quast C, Schweer T, Peplies J, Ludwig W, Glöckner FO (2014) The SILVA and ‘all-species living tree project (LTP)’ taxonomic frameworks. Nucleic Acids Research 42, D643–D648.
The SILVA and ‘all-species living tree project (LTP)’ taxonomic frameworks.Crossref | GoogleScholarGoogle Scholar | 24293649PubMed |

Zened A, Combes S, Cauquil L, Mariette J, Klopp C, Bouchez O, Troegeler-Meynadier A, Enjalbert F (2013) Microbial ecology of the rumen evaluated by 454 GS FLX pyrosequencing is affected by starch and oil supplementation of diets. FEMS Microbiology Ecology 83, 504–514.
Microbial ecology of the rumen evaluated by 454 GS FLX pyrosequencing is affected by starch and oil supplementation of diets.Crossref | GoogleScholarGoogle Scholar | 22974422PubMed |