Effects of Moringa oleifera leaf extract on ruminal methane and carbon dioxide production and fermentation kinetics in a steer model

  • Armando Parra-Garcia
  • Mona Mohamed Mohamed Yasseen Elghandour
  • Ralf Greiner
  • Alberto Barbabosa-Pliego
  • Luis Miguel Camacho-Diaz
  • Abdelfattah Zeidan Mohamed SalemEmail author
Research Article


Ruminal fermentation produces greenhouse gases involved in global warming. Therefore, the effect of nutrient combinations on methane, carbon dioxide, and biogas production as well as ruminal fermentation kinetics was evaluated in in vitro studies. In total mixed rations, dietary corn grain was partially replaced by two levels of soybean hulls (a highly reusable residue), and a Moringa oleifera extract (a natural extract) at three concentration levels was added. Higher levels of both soybean hulls and M. oleifera extract delayed the initiation of methane production and resulted in a lower methane and carbon dioxide production. Thus, total biogas production was also lower. Replacement of corn grain by soybean hulls tended to lower methane production rates and asymptotic carbon dioxide production, and a delay in biogas and methane formation was observed. Asymptotic biogas and carbon dioxide production, however, were increased. The presence of M. oleifera extract tended to delay methane formation and to decrease methane production rate as well as asymptotic methane production. Higher M. oleifera extract levels decreased asymptotic biogas production with the control and the highest soybean hull levels. In the presence of M. oleifera extract, asymptotic carbon dioxide production was shown to be quadratically increased with the control and lowest soybean hull levels, but quadratically decreased with the highest soybean hull level. With the exception of fermentation pH, the interaction of substrate type and M. oleifera extract level was shown to have an effect on all fermentation parameters. Most fermentation parameters were shown to be higher when replacing corn grain by soybean hulls, including fermentation pH. Thus, the conclusion could be drawn that corn grain replacement by soybean hulls (an agricultural residue) in the presence of M. oleifera extract (a sparing leaf product) could ameliorate greenhouse gas emissions and improve digestion.


Biogas Moringa oleifera extract Ruminal fermentation Soybean hulls 



  1. AOAC (1997) Association of Official Analytical Chemists. Official methods of analysis, 16th ed. AOAC, ArlingtonGoogle Scholar
  2. Blümmel M, Ørskov ER (1993) Comparison of in vitro gas production and nylon bag degradability of roughages in predicting feed intake in cattle. Anim Feed Sci Technol 40:109–119CrossRefGoogle Scholar
  3. Blümmel M, Steingass H, Becker K (1997) The relationship between in vitro gas production, in vitro microbial biomass yield and 15N incorporation and its implications for the prediction of voluntary feed intake of roughages. Br J Nutr 77:911–921CrossRefGoogle Scholar
  4. Blümmel M, Mgomezulu R, Chen XB, Makkar HP, Becker K, Ørskov ER (1999) The modification of an in vitro gas production test to detect roughage related differences in in-vivo microbial protein synthesis as estimated by the excretion of purine derivatives. J Agr Sci Cambridge 133:335–340CrossRefGoogle Scholar
  5. Boadi D, Benchaar C, Chiquette J, Massé D (2004) Mitigation strategies to reduce enteric methane emissions from dairy cows: update review. Can J Anim Sci 84:319–335CrossRefGoogle Scholar
  6. 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. Anim Feed Sci Technol 176:78–93CrossRefGoogle Scholar
  7. Carulla JE, Kreuzer M, Machmüller A, Hess HD (2005) Supplementation of Acacia mearnsii tannins decreases methanogenesis and urinary nitrogen in forage-fed sheep. Crop Pasture Sci 56:961–970CrossRefGoogle Scholar
  8. Cedillo J, Vázquez-Armijo JF, González-Reyna A, Salem AZ, Kholif AE, Hernández-Meléndez J, Martínez-González JC, Jiménez RMDO, Rivero N, López D (2014) Effects of different doses of Salix babylonica extract on growth performance and diet in vitro gas production in Pelibuey growing lambs. Ital J Anim Sci 13:609–613CrossRefGoogle Scholar
  9. Costa SBM, Ferreira MA, Pessoa RAS, Batista AMV, Ramos AO, da Conceição MG, Gomes LHS (2012) Tifton hay, soybean hulls, and whole cottonseed as fiber source in spineless cactus diets for sheep. Trop Anim Health Prod 44:1993–2000CrossRefGoogle Scholar
  10. Dey A, Paul SS, Pandey P, Rathore R (2014) Potential of Moringa oleifera leaves in modulating in vitro methanogenesis and fermentation of wheat straw in buffalo. Indian J Anim Sci 84:533–538Google Scholar
  11. Elghandour MMY, Kholif AE, Marquez-Molina O, Vazquez-Armijo JF, Puniya AK, Salem AZM (2015) Influence of individual or mixed cellulase and xylanase mixture on in vitro rumen gas production kinetics of total mixed rations with different maize silage and concentrate ratios. Turk J Vet Anim Sci 39:435–442CrossRefGoogle Scholar
  12. Elghandour MMY, Kholif AE, Salem AZM, Olafadehan OA, Kholif AM (2016) Sustainable anaerobic rumen methane and carbon dioxide productions from prickly pear cactus flour by organic acid salts addition. J Clean Prod 139:1362–1369Google Scholar
  13. FAO (Food and Agriculture Organization of the United Nations) (2006) Livestock a major threat to the environment: remedies urgently needed. [Accessed 15 August 2016]. Available from
  14. Fenchel T, Finlay BJ (1995) Oxford series in ecology and evolution. Oxford University Press, Ecology and evolution in anoxic worlds.Google Scholar
  15. France J, Dijkstra J, Dhanoa MS, Lopez S, Bannink A (2000) Estimating the extent of degradation of ruminant feeds from a description of their gas production profiles observed in vitro: derivation of models and other mathematical considerations. Br J Nutr 83:143–150CrossRefGoogle Scholar
  16. Getachew G, Makkar HPS, Becker K (2002) Tropical browses: contents of phenolic compounds, in vitro gas production and stoichiometric relationship between short chain fatty acid and in vitro gas production. J Agr Sci Cambridge 139:341–352Google Scholar
  17. Goering MK, van Soest PJ (1970) Forage fiber analysis (apparatus, reagents, procedures and some applications). Agriculture handbook, no 379. Agricultural Research Service, USDA, Washington, DCGoogle Scholar
  18. Hart KJ, Yanez-Ruiz DR, Duval SM, McEwan NR, Newbold CJ (2008) Plant extracts to manipulate rumen fermentation. Anim Feed Sci Technol 147(1):8–35CrossRefGoogle Scholar
  19. Hook SE, Wright ADG, McBride BW (2010) Methanogens: methane producers of the rumen and mitigation strategies. Archaea 945785.
  20. Iqbal MF, Cheng YF, Zhu WY, Zeshan B (2008) Mitigation of ruminant methane production: current strategies, constraints and future options. World J Microbiol Biotechnol 24:2747–2755CrossRefGoogle Scholar
  21. Johnson KA, Johnson DE (1995) Methane emissions from cattle. J Anim Sci 73:2483–2492CrossRefGoogle Scholar
  22. Khazaal KA, Parissi Z, Tsiouvaras C, Nastis A, Ørskov ER (1996) Assessment of phenolics-related antinutritive levels using the in vitro gas production technique: a comparison between different types of polyvinylpyrrolidone or polyethylene glycol. J Sci Food Agric 71:405–414CrossRefGoogle Scholar
  23. Kholif AE, Gouda GA, Morsy TA, Salem AZM, Lopez S, Kholif AM (2015) Moringa oleifera leaf meal as a protein source in lactating goat’s diets: feed intake, digestibility, ruminal fermentation, milk yield and composition, and its fatty acids profile. Small Rumin Res 129:129–137CrossRefGoogle Scholar
  24. Kholif A, Elghandour M, Salem A, Barbabosa A, Márquez O, Odongo N (2017) The effects of three total mixed rations with different concentrate to maize silage ratios and different levels of microalgae Chlorella vulgaris on in vitro total gas, methane and carbon dioxide production. J Agric Sci 155(3):494–507CrossRefGoogle Scholar
  25. Menke KH, Raab L, Salewski A, Steingass H, Fritz D, Schneider W (1979) 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 Agr Sci Cambridge 93:217–222CrossRefGoogle Scholar
  26. Mueller-Harvey I (2006) Review, unraveling the conundrum of tannins in animal nutrition and health. J Sci Food Agric 86:2010–2037CrossRefGoogle Scholar
  27. NRC (2001) Nutrient requirement of dairy cattle, 7th rev. edn. National Research Council, National Academy Press, Washington, DCGoogle Scholar
  28. Ørskov ER, Ryle R (1990) Energy nutrition in ruminants. Elsevier, New YorkGoogle Scholar
  29. Polyorach S, Wanapat M, Cherdthong A (2014) Influence of yeast fermented cassava chip protein (YEFECAP) and roughage to concentrate ratio on ruminal fermentation and microorganisms using in vitro gas production technique. Asian Australas J Anim Sci 27:36–45CrossRefGoogle Scholar
  30. Ramos S, Tejido ML, Martínez ME, Ranilla MJ, Carro MD (2009) Microbial protein synthesis, ruminal digestion, microbial populations, and nitrogen balance in sheep fed diets varying in forage-to-concentrate ratio and type of forage. J Anim Sci 87:2924–2934CrossRefGoogle Scholar
  31. Salem AZM, Szumacher-Strabel M, Lopez S, Khalil MS, Mendoza GD, Ammar H (2012) In situ degradability of soyabean meal treated with Acacia saligna and Atriplex halimus extracts in sheep. J Anim Feed Sci 21(3):447–457CrossRefGoogle Scholar
  32. Salem AZM, Kholif AE, Elghandour MMY, Hernandez SR, Domínguez-Vara IA, Mellado M (2014) Effect of increasing levels of seven tree species extracts added to a high concentrate diet on in vitro rumen gas output. Anim Sci J 85(9):853–860CrossRefGoogle Scholar
  33. SAS (2002) Statistical Analysis System. User’s guide: statistics. Ver 9.0. SAS Institute, CaryGoogle Scholar
  34. Sirohi SK, Pandey N, Singh B, Puniya AK (2010) Rumen methanogens: a review. Indian J Microbiol 50:253–262CrossRefGoogle Scholar
  35. Soliva CR, Kreuzer M, Foidl N, Foidl G, Machmuller A, Hess HD (2005) Feeding value of whole and extracted Moringa oleifera leaves for ruminants and their effects on ruminal fermentation in vitro. Anim Feed Sci Technol 118:47–62CrossRefGoogle Scholar
  36. Theodorou MK, Williams BA, Dhanoa MS, McAllan AB, France J (1994) A simple gas production method using a pressure transducer to determine the fermentation kinetics of ruminant feeds. Anim Feed Sci Technol 48:185–197CrossRefGoogle Scholar
  37. Tiemann TT, Lascano CE, Wettstein H-R, Mayer AC, Kreuzer M, Hess HD (2008) Effect of the tropical tanninrich shrub legumes Calliandra calothyrsus and Flemingia macrophylla on methane emission and nitrogen and energy balance in growing lambs. Animal 2:790–799CrossRefGoogle Scholar
  38. Van Soest PJ, Robertson JB, Lewis BA (1991) Methods for dietary fibre, neutral detergent fibre, and non-starch carbohydrates in relation to animal nutrition. J Dairy Sci 74:3583–3597CrossRefGoogle Scholar
  39. Walsh K, O’Kiely P, Taweel H, McGee M, Moloney AP, Boland TM (2009) Intake, digestibility and rumen characteristics in cattle offered whole-crop wheat or barley silages of contrasting grain to straw ratios. Anim Feed Sci Technol 148:192–213CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Armando Parra-Garcia
    • 1
  • Mona Mohamed Mohamed Yasseen Elghandour
    • 1
  • Ralf Greiner
    • 2
  • Alberto Barbabosa-Pliego
    • 1
  • Luis Miguel Camacho-Diaz
    • 3
  • Abdelfattah Zeidan Mohamed Salem
    • 1
    Email author
  1. 1.Facultad de Medicina Veterinaria y ZootecniaUniversidad Autónoma del Estado de MéxicoTolucaMexico
  2. 2.Department of Food Technology and Bioprocess Engineering, Federal Research Institute of Food and NutritionMax Rubner-InstitutKarlsruheGermany
  3. 3.Facultad de Medicina Veterinaria y ZootecniaUniversidad Autónoma de GuerreroCiudad Altamirano, GuerreroMexico

Personalised recommendations