In vitro five brown algae extracts for efficiency of ruminal fermentation and methane yield


This study aimed to evaluate the effects of extracts of five species of brown algae (Ecklonia stolonifera, ESA; Eisenia bicyclis, EBS; Sargarssum fulvellum, SFM; Undaria pinnatifida, UPA; Sargassum fusiforme, SFS) on in vitro ruminal fermentation characteristics, total gas and methane production, and rumen microbial populations when incubated with grass (timothy, Phleum pratense) as the primary substrate. Rumen fluid donors were two rumen-fistulated Holstein cows with free access to water and mineralized salt block. An in vitro trial was carried out using 6, 12, 24, 48, and 72 h incubation with brown algae extracts added at concentration of 5% of timothy. Digestibility of dry matter (DM) was highest for SFS compared with control (Ctrl), with the remaining treatments being intermediate and similar. Ammonia nitrogen concentration was significantly higher at ESA and SFS than Ctrl. The concentrations of total VFA, acetate, and propionate were higher in all treatments compared with Ctrl, except for the propionate concentration at 48 h incubation. Total gas production of all treatments significantly increased with incubation time compared with Ctrl, whereas methane production was significantly decreased after 48 h incubation. As determined by relative quantification of specific ruminal microbes, brown algae extracts significantly affected the abundance of cellulolytic bacteria (i.e., Ruminococcus albus, Fibrobacter succinogenes, Ruminococcus flavefaciens), methanogenic archaea, and ciliate-associated methanogens. These results suggest that supplementation of brown algae extracts can modify ruminal fermentation to increase VFA concentration and total gas production and alter ammonia nitrogen and methane production.

This is a preview of subscription content, access via your institution.

Fig. 1


  1. Alexander G, Singh B, Sahoo A, Bhat TK (2008) In vitro screening of plant extracts to enhance the efficiency of utilization of energy and nitrogen in ruminant diets. Anim Feed Sci Technol 145:229–244

    CAS  Article  Google Scholar 

  2. AOAC (1990) Official Methods of Analysis, 15th edn. Assoc. of Official Anal. Chem, Arlington

    Google Scholar 

  3. Bhatta R, Tajima K, Kurihara M (2006) Influence of temperature and pH on fermentation pattern and methane production in the rumen simulating fermenter (RUSITEC). Asian Australas J Anim Sci 19:376–380

    CAS  Article  Google Scholar 

  4. Božic AK, Anderson RC, Carstens GE, Ricke SC, Callaway TR, Yokoyama MT, Wand JK, Nisbet DJ (2009) Effects of the methane-inhibitors nitrate, nitroethane, lauric acid, Lauricidin® and the Hawaiian marine algae Chaetoceros on ruminal fermentation in vitro. Bioresour Technol 100:4017–4025

  5. Bryukhanov AL, Netrusov AI (2004) Review: Catalase catalase and superoxide dismutase: Distributiondistribution, properties, and physiological role in cells of strict anaerobes. Biokhimiya 69:1170–1186

    Google Scholar 

  6. Chaney AL, Marbach EP (1962) Modified reagents for determination of urea and ammonia. Clin Chem 8:130–132

    CAS  Article  Google Scholar 

  7. Choi YY, Lee SJ, Kim HS, EOM JS, Kim DH, Lee SS (2020a) The potential nutritive value of Sargassum fulvellum as a feed ingredient for ruminants. Algal Res 45:101761

  8. Choi YY, Lee SJ, Lee YJ, Kim HS, Eom JS, Jo SU, Lee SS (2020b) In vitro and in situ evaluation of Undaria pinnatifida as a feed ingredient for ruminants. J Appl Phycol 32:729–739

  9. Choi YY, Lee SJ, Lee YJ, Kim HS, Eom JS, Kim SC, Kim ET, Lee SS (2020c) New challenges for efficient usage of Sargassum fusiforme for ruminant production. Sci Rep 10:1–13

  10. Denman SE, McSweeney CS (2006) Development of a real-time PCR assay for monitoring anaerobic fungal and cellulolytic bacterial populations within the rumen. FEMS Microbiol Ecol 58:572–582

    CAS  Article  Google Scholar 

  11. Denman SE, Tomkins NW, McSweeney CS (2007) Quantitation and diversity analysis of ruminal methanogenic populations in response to the antimethanogenic compound bromochloromethane. FEMS Microbiol Ecol 62:313–322

    CAS  Article  Google Scholar 

  12. Gerber PJ, Steinfeld H, Henderson B, Mottet A, Opio C, Dijkman J, Falucci A, Tempio G (2013) Tackling climate change through livestock: a global assessment of emissions and mitigation opportunities. Food and Agriculture Organization of the United Nations (FAO), Rome

    Google Scholar 

  13. Getachew G, Robinson PH, DePeters EJ, Taylor SJ (2004) Relationships between chemical composition, dry matter degradation and in vitro gas production of several ruminant feeds. Anim Feed Sci Technol 111:57–71

    CAS  Article  Google Scholar 

  14. Hristov AN, Oh J, Firkins JL, Dijkstra J, Kebreab E, Waghorn G, Makkar HPS, Adesogan AT, Yang W, Lee C, Gerber PJ, Henderson B, Tricarico JM (2013) Special topics—Mitigation mitigation of methane and nitrous oxide emissions from animal operations: I. A review of enteric methane mitigation options. J Anim Sci 91:5045–5069

    CAS  Article  Google Scholar 

  15. Hwang HS, Ok JU, Lee SJ, Chu GM, Kim KH, Oh YK, Lee SS, Lee SS (2012) Effects of halogenated compounds on in vitro fermentation characteristics in the rumen and methane emissions. J Life Sci 22:1187–1193 (In Korean)

    Article  Google Scholar 

  16. Kim ET, Hwang HS, Lee SM, Lee SJ, Lee ID, Lee SK, Oh da S, Lim JH, Yoon HB, Jeong HY, Im SK, Lee SS (2016a) Effects of medicinal herb extracts on in vitro ruminal methanogenesis, microbe diversity and fermentation system. Asian Australas J Anim Sci 29:1280–1286

    Article  Google Scholar 

  17. Kim SH, Mamuad LL, Kim DW, KIm SK, Lee SS (2016b) Fumarate reductase-producing enterococci reduce methane production in rumen fermentation in vitro. J Microbiol Biotechnol 26:558–566

    CAS  Article  Google Scholar 

  18. Koike S, Kobayashi Y (2001) Development and use of competitive PCR assays for the rumen cellulolytic bacteria: Fibrobacter succinogenes, Ruminococcus albus and Ruminococcus flavefaciens. FEMS Microbiol Lett 204:361–366

  19. Kumar S, Puniya AK, Puniya M, Dagar SS, Sirohi SK, Singh K, Griffith GW (2009) Factors affecting rumen methanogens and methane mitigation strategies. World J Microbiol Biotechnol 25:1557–1566

    Article  Google Scholar 

  20. Lee JH, Kim GH (2015) Evaluation of antioxidant activity of marine algae-extracts from Korea. J Aquat Food Prod Technol 24:227–240

    CAS  Article  Google Scholar 

  21. Lee JW, Seok JK, Boo YC (2018) Ecklonia cava extract and dieckol attenuate cellular lipid peroxidation in keratinocytes exposed to PM10. Evid Based Complement Alternat Med 2018:8248323

    PubMed  PubMed Central  Google Scholar 

  22. Machado L, Magnusson M, Paul NA, de Nys R, Tomkins N (2014) Effects of marine and freshwater macroalgae on in vitro total gas and methane production. PLoS One 9:e0085289

    Google Scholar 

  23. Ørskov ER, McDonald I (1979) The estimation of protein degradability in the rumen from incubation measurements weighted according to rate of passage. J Agric Sci 92:499–503

    Article  Google Scholar 

  24. Patra AK, Yu Z (2012) Effects of essential oils on methane production and fermentation by, and abundance and diversity of, rumen microbial populations. Appl Environ Microbiol 78:4271–4280

    CAS  Article  Google Scholar 

  25. Patra AK, Stiverson J, Yu Z (2012) Effects of quillaja and yucca saponins on communities and select populations of rumen bacteria and archaea, and fermentation in vitro. J Appl Microbiol 113:1329–1340

    CAS  Article  Google Scholar 

  26. Shingu H, Hayashi H, Touno E, Oshibe A, Kushibiki S, Oda S, Katoh K, Obara Y (2007) Characteristics of developmental changes in the kinetics of glucose and urea in Japanese Black calves: Comparison comparison with Holstein calves. J Anim Sci 85:2910–2915

    CAS  Article  Google Scholar 

  27. Skillman LC, Skillman LC, Toovey AF, Williams AJ, Wright AG (2006) Development and validation of a real-time PCR method to quantify rumen protozoa and examination of variability between. Cloning 72:200–206

    CAS  Google Scholar 

  28. Stocker TF, Qin D, Plattner GK, Tignor M, Allen SK, Boschung J, Nauels A, Xia Y, Bex V, Midgley PM (eds) (2013) Climate Change 2013. The Physical Science Basis. Working Group I Contribution to the Fifth Assessment Report of the Intergovernmental Panel on Climate Change. Cambridge University Press, Cambridge, p 1535

    Google Scholar 

  29. Swanson AK, Druehl LD (2002) Induction, exudation and the UV protective role of kelp phlorotannins. Aquat Bot 73:241–253

    CAS  Article  Google Scholar 

  30. 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 48:185–197

    Article  Google Scholar 

  31. Van Nevel CJ (1996) Control of rumen methanogenesis. Environ Monit Assess 42:73–97

    Article  Google Scholar 

  32. Vissers AM, Pellikaan WF, Bouwhuis A, Vincken J-P, Gruppen H, Hendriks WH (2018) Laminaria digitata phlorotannins decrease protein degradation and methanogenesis during in vitro ruminal fermentation. J Sci Food Agric 98:3644–3650

  33. Wang Y, Alexander TW, Mcallister TA (2009) In vitro effects of phlorotannins from Ascophyllum nodosum (brown seaweed) on rumen bacterial populations and fermentation. J Sci Food Agric 89:2252–2260

  34. Watanabe Y, Suzuki R, Koike S, Nagashima K, Mochizuki M, Forster RJ, Kobayashi Y (2010) In vitro evaluation of cashew nut shell liquid as a methane-inhibiting and propionate-enhancing agent for ruminants. J Dairy Sci 93:5258–5267

    CAS  Article  Google Scholar 

  35. Williams AG, Withers S, Sutherland AD (2013) The potential of bacteria isolated from ruminal contents of seaweed-eating North Ronaldsay sheep to hydrolyse seaweed components and produce methane by anaerobic digestion in vitro. Microb Biotechnol 6:45–52

    Article  Google Scholar 

  36. Wu W, Hasumi K, Peng H, Hu X, Wang X, Bao B (2009) Fibrinolytic compounds isolated from a brown alga, Sargassum fulvellum. Mar Drugs 7:85–94

  37. Zheng Y, Xue S, Zhao Y, Li S (2020) Effect of Cassava Residue Substituting for Crushed Maize on In Vitro Ruminal Fermentation Characteristics of Dairy Cows at Mid-Lactation. Animals 10(5):893

    Article  Google Scholar 

Download references


This work was supported by the National Research Foundation (NRF) of Korea Grant funded by the Korean Government (grant number NRF-2015R1A6A1A03031413).

Author information



Corresponding author

Correspondence to Sung Sill Lee.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Choi, Y.Y., Shin, N.H., Lee, S.J. et al. In vitro five brown algae extracts for efficiency of ruminal fermentation and methane yield. J Appl Phycol 33, 1253–1262 (2021).

Download citation


  • Phaeophyceae
  • Methane
  • Rumen microorganism
  • Ruminant nutrition
  • Seaweed