Register      Login
Animal Production Science Animal Production Science Society
Food, fibre and pharmaceuticals from animals
RESEARCH ARTICLE

Essential oils from Lippia turbinata and Tagetes minuta persistently reduce in vitro ruminal methane production in a continuous-culture system

F. Garcia A B F , P. E. Vercoe C , M. J. Martínez D , Z. Durmic C , M. A. Brunetti D , M. V. Moreno D , D. Colombatto A E , E. Lucini B and J. Martínez Ferrer D
+ Author Affiliations
- Author Affiliations

A Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Godoy Cruz 2290, Buenos Aires, C1425FQB, Argentina.

B Facultad de Ciencias Agropecuarias, Universidad Nacional de Córdoba, Félix Aldo Marrone 746, Córdoba, 5000, Argentina.

C School of Agriculture and Environment, University of Western Australia, 35 Stirling Highway, Crawley, WA 6009, Australia.

D Instituto Nacional de Tecnología Agropecuaria, Ruta Nacional 9, Km 636, Manfredi, 5988, Argentina.

E Facultad de Agronomía, Universidad de Buenos Aires, Avenida San Martín 4453, Buenos Aires, C1417DSQ, Argentina.

F Corresponding author. Email: fgarcia@agro.unc.edu.ar

Animal Production Science 59(4) 709-720 https://doi.org/10.1071/AN17469
Submitted: 13 July 2017  Accepted: 11 February 2018   Published: 24 May 2018

Abstract

The aim of the present study was to evaluate the impact of essential oils (EO) from Lippia turbinata (LT) and Tagetes minuta (TM) as well as the rotation of both EO on fermentation parameters in vitro. Daily addition of LT, TM, or a 3-day rotation between them (TM/LT), as well as a control (without EO), was evaluated using the rumen simulation technique (Rusitec). The experiment lasted 19 days, with a 7-day adaptation period, followed by 12 days of treatment (Days 0–12). The EO were dissolved in ethanol (70% vol/vol) to be added daily to fermenters (300 μL/L) from Day 0. Daily measurements included methane concentration, total gas production, apparent DM disappearance and pH, which started 2 days before the addition of treatments. On Days 0, 4, 8 and 12 apparent crude protein disappearance and neutral detergent fibre disappearance, ammonia and volatile fatty acid concentration and composition were determined. Methane production was significantly inhibited shortly after addition of both EO added individually, and persisted over time with no apparent adaptation to EO addition. The TM/LT treatment showed a similar effect on methane production, suggesting that rotating the EO did not bring further improvements in reduction or persistency compared with the inclusion of the EO individually. Gas production, total volatile fatty acid concentration and composition and apparent crude protein disappearance were not affected by EO addition. Compared with the control, a 5% reduction of apparent DM disappearance and a 15% reduction of neutral detergent fibre disappearance were observed with the addition of EO. Only TM and TM/LT reduced ammonia concentration. Given the significant and persistent antimethanogenic activity of both EO, and the potential of T. minuta to modify nitrogen metabolism, EO from these plant species are of interest for developing new feed additives with potential application in ruminant nutrition that are also likely to be acceptable to consumers.

Additional keywords: adaptation, greenhouse gases, supplements, ruminants, Rusitec, semi-continuous cultures.


References

Acamovic T, Brooker JD (2005) Biochemistry of plant secondary metabolites and their effects in animals. Proceedings of the Nutrition Society 64, 403–412.

Adams RP (1995) ‘Identification of essential oil components by gas chromatography/mass spectrometry.’ (Allured Publishing Corporation: Carol Stream, IL)

AOCS (1998) ‘Official methods and recommended practices of the American Oil Chemists’ Society.’ 5th edn. (American Oil Chemist Society: Champaign, IL)

Bakkali F, Averbeck S, Averbeck D, Idaomar M (2008) Biological effects of essential oils: a review. Food and Chemical Toxicology 46, 446–475.
Biological effects of essential oils: a review.Crossref | GoogleScholarGoogle Scholar |

Banik BK, Durmic Z, Erskine W, Revell CK, Vadhanabhuti J, McSweeney CS, Padmanabha J, Flematti GR, Algreiby AA, Vercoe PE (2016) Bioactive fractions from the pasture legume Biserrula pelecinus L. have an anti-methanogenic effect against key rumen methanogens. Anaerobe 39, 173–182.
Bioactive fractions from the pasture legume Biserrula pelecinus L. have an anti-methanogenic effect against key rumen methanogens.Crossref | GoogleScholarGoogle Scholar |

Benchaar C, Greathead H (2011) Essential oils and opportunities to mitigate enteric methane emissions from ruminants. Animal Feed Science and Technology 166–167, 338–355.
Essential oils and opportunities to mitigate enteric methane emissions from ruminants.Crossref | GoogleScholarGoogle Scholar |

Bergmeyer HU, Beutler HO (1985) Ammonia. In ‘Methods of enzymatic analysis. Vol. 8’. 3rd edn. (Ed. HU Bergmeyer) pp. 454–461. (Academic Press: New York)

Bodas R, Prieto N, García-González R, Andrés S, Giráldez FJ, López S (2012) Manipulation of rumen fermentation and methane production with plant secondary metabolites. Animal Feed Science and Technology 176, 78–93.
Manipulation of rumen fermentation and methane production with plant secondary metabolites.Crossref | GoogleScholarGoogle Scholar |

Burt S (2004) Essential oils: their antibacterial properties and potential applications in foods. A review. International Journal of Food Microbiology 94, 223–253.
Essential oils: their antibacterial properties and potential applications in foods. A review.Crossref | GoogleScholarGoogle Scholar |

Busquet M, Calsamiglia S, Ferret A, Kamel C (2005a) Screening for effects of plant extracts and active compounds of plants on dairy cattle rumen microbial fermentation in a continuous culture system. Animal Feed Science and Technology 123–124, 597–613.
Screening for effects of plant extracts and active compounds of plants on dairy cattle rumen microbial fermentation in a continuous culture system.Crossref | GoogleScholarGoogle Scholar |

Busquet M, Calsamiglia S, Ferret A, Cardozo PW, Kamel C (2005b) Effects of cinnamaldehyde and garlic oil on rumen microbial fermentation in a dual flow continuous culture. Journal of Dairy Science 88, 2508–2516.
Effects of cinnamaldehyde and garlic oil on rumen microbial fermentation in a dual flow continuous culture.Crossref | GoogleScholarGoogle Scholar |

Busquet M, Calsamiglia S, Ferret A, Kamel C (2006) Plant extracts affect in vitro rumen microbial fermentation. Journal of Dairy Science 89, 761–771.
Plant extracts affect in vitro rumen microbial fermentation.Crossref | GoogleScholarGoogle Scholar |

Cardozo PW, Calsamiglia S, Ferret A, Kamel C (2004) Effects of natural plant extracts on ruminal protein degradation and fermentation profiles in continuous culture. Journal of Animal Science 82, 3230–3236.
Effects of natural plant extracts on ruminal protein degradation and fermentation profiles in continuous culture.Crossref | GoogleScholarGoogle Scholar |

Castillejos L (2005) Modificación de la fermentación microbiana ruminal mediante compuestos de aceites esenciales. PhD Thesis, Universitat Autònoma de Barcelona, Spain.

Castillejos L, Calsamiglia S, Ferret A (2006) Effect of essential oil active compounds on rumen microbial fermentation and nutrient flow in in vitro systems. Journal of Dairy Science 89, 2649–2658.
Effect of essential oil active compounds on rumen microbial fermentation and nutrient flow in in vitro systems.Crossref | GoogleScholarGoogle Scholar |

Cattani M, Maccarana L, Rossi G, Tagliapietra F, Schiavon S, Bailoni L (2016) Dose-response and inclusion effects of pure natural extracts and synthetic compounds on in vitro methane production. Animal Feed Science and Technology 218, 100–109.
Dose-response and inclusion effects of pure natural extracts and synthetic compounds on in vitro methane production.Crossref | GoogleScholarGoogle Scholar |

Chamorro ER, Ballerini G, Sequeira AF, Velasco GA, Zalazar MF (2008) Chemical composition of essential oil from Tagetes minuta L. leaves and flowers. The Journal of Argentine Chemical Society 96, 80–86.

Chizzola R, Hochsteiner W, Hajek S (2004) GC analysis of essential oils in the rumen fluid after incubation of Thuja orientalis twigs in the Rusitec system. Veterinary Science 76, 77–82.
GC analysis of essential oils in the rumen fluid after incubation of Thuja orientalis twigs in the Rusitec system.Crossref | GoogleScholarGoogle Scholar |

Cobellis G, Trabalza-Marinucci M, Marcotullio MC, Yu Z (2016) Evaluation of different essential oils in modulating methane and ammonia production, rumen fermentation, and rumen bacteria in vitro. Animal Feed Science and Technology 215, 25–36.
Evaluation of different essential oils in modulating methane and ammonia production, rumen fermentation, and rumen bacteria in vitro.Crossref | GoogleScholarGoogle Scholar |

Czerkawski JW, Breckenridge G (1977) Design and development of a long-term rumen simulation technique (Rusitec). British Journal of Nutrition 38, 371–384.
Design and development of a long-term rumen simulation technique (Rusitec).Crossref | GoogleScholarGoogle Scholar |

Dellacasa AD, Bailac PN, Ponzi MI, Ruffinengo SR, Eguaras MJ (2003) In vitro activity of essential oils from San Luis–Argentina against Ascosphaera apis. The Journal of Essential Oil Research 15, 282–285.
In vitro activity of essential oils from San Luis–Argentina against Ascosphaera apis.Crossref | GoogleScholarGoogle Scholar |

Dewick PM (2002) ‘Medicinal natural products: a biosynthetic approach.’ (John Wiley & Sons: Chichester, UK)

Di Rienzo JA, Guzman AW, Casanoves F (2002) A multiple-comparisons method based on the distribution of the root node distance of a binary tree. Journal of Agricultural Biological & Environmental Statistics 7, 129–142.
A multiple-comparisons method based on the distribution of the root node distance of a binary tree.Crossref | GoogleScholarGoogle Scholar |

Di Rienzo JA, Casanoves F, Balzarini MG, Gonzalez L, Tablada M, Robledo CW (2016) ‘InfoStat software.’ Available at http://www.infostat.com.ar/ [Verified January 2017]

Dorman HJD, Deans SG (2000) Antimicrobial agents from plants: antibacterial activity of plant volatile oils. Journal of Applied Microbiology 88, 308–316.
Antimicrobial agents from plants: antibacterial activity of plant volatile oils.Crossref | GoogleScholarGoogle Scholar |

Fraser GR, Chaves AV, Wang Y, McAllister TA, Beauchemin KA, Benchaar C (2007) Assessment of the effects of cinnamon leaf oil on rumen microbial fermentation using two continuous culture systems. Journal of Dairy Science 90, 2315–2328.
Assessment of the effects of cinnamon leaf oil on rumen microbial fermentation using two continuous culture systems.Crossref | GoogleScholarGoogle Scholar |

García CC, Talarico L, Almeida N, Colombres S, Duschatzky C, Damonte EB (2003) Virucidal activity of essential oils from aromatic plants of San Luis, Argentina. Phytotherapy Research 17, 1073–1075.
Virucidal activity of essential oils from aromatic plants of San Luis, Argentina.Crossref | GoogleScholarGoogle Scholar |

Garcia MV, Matias J, Cavalcante Barros JC, Pries de Lima D, da Silva Lopes R, Andreotti R (2012) Chemical identification of Tagetes minuta Linnaeus (Asteraceae) essential oil and its acaricidal effect on ticks. Revista Brasileira de Parasitologia Veterinária 21, 405–411.
Chemical identification of Tagetes minuta Linnaeus (Asteraceae) essential oil and its acaricidal effect on ticks.Crossref | GoogleScholarGoogle Scholar |

Garcia F, Martínez Ferrer J, Durmic Z, Vercoe PE (2015) Effect of ethanol as carrier of essential oils on in vitro methane production. In ‘XXIV reunión de la Asociación Latinoamericana de Producción Animal’, 9–13 November 2015, Puerto Varas, Chile. p. 518.

Garcia F, Martinez Ferrer J, Cora A, Brunetti MA, Frossasco G, Moreno MV, Lucini E, Martínez MJ, Colombatto D (2016) Effect of essential oils from Argentinian native species on in vitro methane production. In ‘Program book of the 6th greenhouse gas and animal agriculture conference’, 14–18 February 2016, Melbourne, Australia. p. 83.

Héthélyi E, Dános B, Tétényi P (1986) GC-MS Analysis of the essential oils of four Tagetes species and the anti-microbial activity of Tagetes minuta. Flavour and Fragrance Journal 1, 169–173.
GC-MS Analysis of the essential oils of four Tagetes species and the anti-microbial activity of Tagetes minuta.Crossref | GoogleScholarGoogle Scholar |

IPCC (2013) Summary for policymakers, technical summary and frequently asked questions. In ‘Climate change 2013: the physical science basis’. Contribution of Working Group I to the Fifth Assessment Report of the Intergovernmental Panel on Climate Change. (Eds TF Stocker, D Qin, G-K Plattner, M Tignor, SK Allen, J Boschung, A Nauels, Y Xia, V Bex, PM Midgley) (Cambridge University Press: Cambridge, UK)

Janssen PH (2010) Influence of hydrogen on rumen methane formation and fermentation balances through microbial growth kinetics and fermentation thermodynamics. Animal Feed Science and Technology 160, 1–22.
Influence of hydrogen on rumen methane formation and fermentation balances through microbial growth kinetics and fermentation thermodynamics.Crossref | GoogleScholarGoogle Scholar |

Joch M, Cermak L, Hakl J, Hucko B, Duskova D, Marounek M (2016) In vitro screening of essential oil active compounds for manipulation of rumen fermentation and methane mitigation. Asian–Australasian Journal of Animal Sciences 29, 952–959.
In vitro screening of essential oil active compounds for manipulation of rumen fermentation and methane mitigation.Crossref | GoogleScholarGoogle Scholar |

Johnson MC, Devine AA, Ellis JC, Grunden AM, Fellner V (2009) Effects of antibiotics and oil on microbial profiles and fermentation in mixed cultures of ruminal microorganisms. Journal of Dairy Science 92, 4467–4480.
Effects of antibiotics and oil on microbial profiles and fermentation in mixed cultures of ruminal microorganisms.Crossref | GoogleScholarGoogle Scholar |

Juliani HR, Koroch A, Simon JE, Biurrun FN, Castellano V, Zygadlo JA (2004) Essential oils from Argentinean aromatic plants. Acta Horticulturae 629, 491–498.

Khiaosa-ard R, Zebeli Q (2013) Meta-analysis of the effects of essential oils and their bioactive compounds on rumen fermentation characteristics and feed efficiency in ruminants. Journal of Animal Science 91, 1819–1830.
Meta-analysis of the effects of essential oils and their bioactive compounds on rumen fermentation characteristics and feed efficiency in ruminants.Crossref | GoogleScholarGoogle Scholar |

Klevenhusen F, Muro-Reyes A, Khiaosa-ard R, Metzler-Zebeli BU, Zebeli Q (2012) A meta-analysis of effects of chemical composition of incubated diet and bioactive compounds on in vitro ruminal fermentation. Animal Feed Science and Technology 176, 61–69.
A meta-analysis of effects of chemical composition of incubated diet and bioactive compounds on in vitro ruminal fermentation.Crossref | GoogleScholarGoogle Scholar |

Kouazounde JB, Jin L, McAllister TA, Gbenou JD (2016) In vitro screening of selected essential oils from medicinal plants acclimated to Benin for their effects on methane production from rumen microbial fermentation. African Journal of Biotechnology 15, 442–450.
In vitro screening of selected essential oils from medicinal plants acclimated to Benin for their effects on methane production from rumen microbial fermentation.Crossref | GoogleScholarGoogle Scholar |

Lin B, Wang JH, Lu Y, Liang Q, Liu JX (2013) In vitro rumen fermentation and methane production are influenced by active components of essential oils combined with fumarate. Journal of Animal Physiology and Animal Nutrition 97, 1–9.
In vitro rumen fermentation and methane production are influenced by active components of essential oils combined with fumarate.Crossref | GoogleScholarGoogle Scholar |

Lock AL, Bauman DE (2004) Modifying milk fat composition of dairy cows to enhance fatty acids beneficial to human health. Lipids 39, 1197–1206.
Modifying milk fat composition of dairy cows to enhance fatty acids beneficial to human health.Crossref | GoogleScholarGoogle Scholar |

Macheboeuf D, Morgavi DP, Papon Y, Mousset J-L, Arturo-Schaan M (2008) Dose-response effects of essential oils on in vitro fermentation activity of the rumen microbial population. Animal Feed Science and Technology 145, 335–350.
Dose-response effects of essential oils on in vitro fermentation activity of the rumen microbial population.Crossref | GoogleScholarGoogle Scholar |

Machmüller A, Ossowski DA, Wanner M, Kreuzer M (1998) Potential of various fatty feeds to reduce methane release from rumen fermentation in vitro (Rusitec). Animal Feed Science and Technology 71, 117–130.
Potential of various fatty feeds to reduce methane release from rumen fermentation in vitro (Rusitec).Crossref | GoogleScholarGoogle Scholar |

Martínez ME, Ranilla MJ, Ramos S, Tejido ML, Carro MD (2011) Evolution of fermentation parameters in Rusitec fermenters operated at different dilution rates and concentrate retention times. In ‘Challenging strategies to promote the sheep and goat sector in the current global context’. (Eds MJ Ranilla, MD Carro, H Ben Salem, P Morand-Fehr) pp. 121–126. (CIHEAM, Universidad de León: Zaragoza, Spain)

Martínez Ferrer J, Garcia F, Brunetti MA, Cora A, Frossasco G, Lucini E, Moreno MV, Martínez MJ, Colombatto D (2014) Evaluación in vitro del potencial anti-metanogénico de aceites esenciales extraídos de plantas nativas de Argentina. In ‘Primera conferencia de gases de efecto invernadero en sistemas agropecuarios de Latinoamérica (GALA)’. (Eds M Alfaro, S González, S Hube, C Muñoz, C Pinares-Patiño, E Ungerfeld) pp. 95–96. (Serie Actas INIA: Osorno, Chile)

McDougall EI (1948) Studies on ruminant saliva. 1. The composition and output of sheep’s saliva. The Biochemical Journal 43, 99–109.
Studies on ruminant saliva. 1. The composition and output of sheep’s saliva.Crossref | GoogleScholarGoogle Scholar |

McEwan NR, Graham RC, Wallace RJ, Losa R, Williams P, Newbold CJ (2002) Effect of essential oils on ammonia production by rumen microbes. Reproduction Nutrition Development 42, S65

McGinn SM, Beauchemin KA, Coates T, Colombatto D (2004) Methane emissions from beef cattle: effects of monensin, sunflower oil, enzymes, yeast, and furmaric acid. Journal of Animal Science 82, 3346–3356.
Methane emissions from beef cattle: effects of monensin, sunflower oil, enzymes, yeast, and furmaric acid.Crossref | GoogleScholarGoogle Scholar |

McIntosh FM, Williams P, Losa R, Wallace RJ, Beever DA, Newbold CJ (2003) Effects of essential oils on ruminal microorganisms and their protein metabolism. Applied and Environmental Microbiology 69, 5011–5014.
Effects of essential oils on ruminal microorganisms and their protein metabolism.Crossref | GoogleScholarGoogle Scholar |

Newbold CJ, McIntosh FM, Williams P, Losa R, Wallace RJ (2004) Effects of a specific blend of essential oil compounds on rumen fermentation. Animal Feed Science and Technology 114, 105–112.
Effects of a specific blend of essential oil compounds on rumen fermentation.Crossref | GoogleScholarGoogle Scholar |

Opio C, Gerber P, Mottet A, Falculli A, Tempio G, MacLeod M, Vellinga T, Henderson B, Steinfeld H (2013) ‘Greenhouse gas emissions from ruminant supply chains: a global life cycle assessment.’ (FAO: Rome)

Patra AK, Yu Z (2012) Effects of essential oils on methane production and fermentation by, and abundance and diversity of, rumen microbial populations. Applied and Environmental Microbiology 78, 4271–4280.
Effects of essential oils on methane production and fermentation by, and abundance and diversity of, rumen microbial populations.Crossref | GoogleScholarGoogle Scholar |

Pérez-Zamora CM, Torres C, Aguado MI, Bela AJ, Nuñez MB, Bregni C (2016) Antibacterial activity of essential oils of Aloysia polystachya and Lippia turbinata (Verbenaceae). Boletín Latinoamericano y del Caribe de Plantas Medicinales y Aromáticas 15, 199–205.

Robinson TP, Bu DP, Carrique-Mas J, Fèvre EM, Gilbert M, Grace D, Hay SI, Jiwakanon J, Kakkar M, Kariuki S, Laxminarayan R, Lubroth J, Magnusson U, Thi Ngoc P, Van Boeckel TP, Woolhouse MEJ (2017) Antibiotic resistance: mitigation opportunities in livestock sector development. Animal 11, 1–3.
Antibiotic resistance: mitigation opportunities in livestock sector development.Crossref | GoogleScholarGoogle Scholar |

Senatore F, Napolitano F, Mohamed M-H, Harris PJC, Mnkeni PNS, Henderson J (2004) Antibacterial activity of Tagetes minuta L. (Asteraceae) essential oil with different chemical composition. Flavour and Fragrance Journal 19, 574–578.
Antibacterial activity of Tagetes minuta L. (Asteraceae) essential oil with different chemical composition.Crossref | GoogleScholarGoogle Scholar |

Steinfeld H, Gerber P, Wassenaar T, Castel V, Rosales M, De Haan C (2006) ‘Livestock’s long shadow: environmental issues and options.’ (FAO: Rome, Italy)

Terblanché FC, Kornelius G (1996) Essential oil constituents of the genus Lippia (Verbenaceae): a literature review. The Journal of Essential Oil Research 8, 471–485.
Essential oil constituents of the genus Lippia (Verbenaceae): a literature review.Crossref | GoogleScholarGoogle Scholar |

Ungerfeld EM (2015) Shifts in metabolic hydrogen sinks in the methanogenesis-inhibited ruminal fermentation: a meta-analysis. Frontiers in Microbiology 6, 1–17.

Van Soest PJ, Robertson JB, Lewis BA (1991) Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. Journal of Dairy Science 74, 3583–3597.
Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition.Crossref | GoogleScholarGoogle Scholar |

Veneman JB, Saetnan ER, Clare AJ, Newbold CJ (2016) MitiGate; an online meta-analysis database for quantification of mitigation strategies for enteric methane emissions. The Science of the Total Environment 572, 1166–1174.
MitiGate; an online meta-analysis database for quantification of mitigation strategies for enteric methane emissions.Crossref | GoogleScholarGoogle Scholar |