Bioaugmentation coupled with phytoremediation for the removal of phenolic compounds and color from treated palm oil mill effluent

  • Palist Jarujareet
  • Korakot Nakkanong
  • Ekawan Luepromchai
  • Oramas SuttinunEmail author
Research Article


The potential for coupling bioaugmentation with phytoremediation to simultaneously treat and utilize treated palm oil mill effluent (TPOME) in animal feed production was determined from a reduction in phenolic compounds and color in soil leachates, as well as from an increased yield of pasture grass. Two phenol-degrading bacteria—Methylobacterium sp. NP3 and Acinetobacter sp. PK1—were inoculated into the Brachiaria humidicola rhizosphere before the application of TPOME. A pot study showed that the soil with both grass and inoculated bacteria had the highest dephenolization and decolorization efficiencies, with a maximum capability of removing 70% from 587 mg total phenolic compounds added and 73% from 4438 color units during ten TPOME application cycles. The results corresponded to increases in the number of phenol-degrading bacteria and the grass yield. In a field study, this treatment was able to remove 46% from 21,453 mg total phenolic compounds added, with a maximum color removal efficiency of 52% from 5105 color units, while the uninoculated plots removed about 24–39% and 29–46% of phenolic compounds and color, respectively. The lower treatment performance was probably due to the increased TPOME concentrations. Based on the amounts of phenolic compounds, protein, and crude fiber in the grass biomass, the inoculated TPOME-treated grass had a satisfactory nutritional quality and digestibility for use as animal feed.


Phytoremediation Bioaugmentation Dephenolization Decolorization Phenol-degrading bacteria Palm oil mill effluent 


Funding information

This research was financially supported by the Thailand Research Fund and Taksin Palm (2521) Co., Ltd. through a Research and Researchers for Industries (RRI) grant (contract no. MSD56I0188), as well as the Graduate School Fund, PSU.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11356_2019_6332_MOESM1_ESM.docx (12.3 mb)
ESM 1 (DOCX 12.3 mb)


  1. Afzal M, Yousaf S, Reichenauer TG, Sessitsch A (2012) The inoculation method affects colonization and performance of bacterial inoculant strains in the phytoremediation of soil contaminated with diesel oil. Int J Phytoremediat 14:35–47CrossRefGoogle Scholar
  2. Afzal M, Khan S, Iqbal S, Mirza MS, Khan QM (2013) Inoculation method affects colonization and activity of Burkholderia phytofirmans PsJN during phytoremediation of diesel-contaminated soil. Int Biodeterior Biodegradation 85:331–336CrossRefGoogle Scholar
  3. Ahmad F, Iqbal S, Anwar S, Afzal M, Islam E, Mustafa T, Khan QM (2012) Enhanced remediation of chlorpyrifos from soil using ryegrass (Lollium multiflorum) and chlorpyrifos-degrading bacterium Bacillus pumilus C2A1. J Hazard Mater 237–238:110–115CrossRefGoogle Scholar
  4. AOAC (2000) Official methods of analysis (17th ed.). Association of Official Analytical Chemists International, MarylandGoogle Scholar
  5. AOAC (2005) Official methods of analysis (18th ed.). Association of Official Analytical Chemists International, MarylandGoogle Scholar
  6. APHA (2005) Standard methods for the examination of water and wastewater (21st ed.). American Public Health Association, Washington DCGoogle Scholar
  7. Aylward OF, McDonald BR, Adams SM, Valenzuela A, Schmidt RA, Goodwin LA et al (2013) Comparison of 26 sphingomonad genomes reveals diverse environmental adaptations and biodegradative capabilities. Appl Environ Microbiol 79:3724–3733CrossRefGoogle Scholar
  8. Basha SA, Sarma BK, Singh DP, Annapurna K, Singh UP (2006) Differential methods of inoculation of plant growth-promoting rhizobacteria induce synthesis of phenylalanine ammonia-lyase and phenolic compounds differentially in chickpea. Folia Microbiol 51:463–468CrossRefGoogle Scholar
  9. Bene CD, Pellegrino E, Debolini M, Silvestri N, Bonari E (2013) Short-term and long-term effects of olive mill wastewater land spreading on soil chemical and biological properties. Soil Biol Biochem 56:21–30CrossRefGoogle Scholar
  10. Bodini SF, Cicalini AR, Santori F (2011) Rhizosphere dynamics during phytoremediation of olive mill wastewater. Bioresour Technol 102:4383–4389CrossRefGoogle Scholar
  11. Canarini A, Kaiser C, Merchant A, Richter A, Wanek W (2019) Root exudation of primary metabolites: mechanisms and their roles in plant responses to environmental stimuli. Front Plant Sci 10:157CrossRefGoogle Scholar
  12. Chantho P, Musikavong C, Suttinun O (2016) Removal of phenolic compounds from palm oil mill effluent by thermophilic Bacillus thermoleovorans strain A2 and their effect on anaerobic digestion. Int Biodeterior Biodegradation 115:293–301CrossRefGoogle Scholar
  13. Chesson A, Stewart CS, Wallace RJ (1982) Influence of plant phenolic acids on growth and cellulolytic activity of rumen bacteria. Appl Environ Microbiol 44:597–603Google Scholar
  14. Coniglio MS, Busto VD, Gonzáles PS, Medina MI, Milrad S, Agostini E (2008) Application of Brassica napus hairy root cultures for phenol removal from aqueous solutions. Chemosphere 72:1035–1042CrossRefGoogle Scholar
  15. Dams RI, Paton G, Killham K (2007) Bioaugmentation of pentachlorophenol in soil and hydroponic system. Int Biodeterior Biodegradation 60:171–177CrossRefGoogle Scholar
  16. Dan A, Fujii D, Soda S, Machimura T, Ike M (2017) Removal of phenol, bisphenol A, and 4-tert-butylphenol from synthetic landfill leachate by vertical flow constructed wetlands. Sci Total Environ 578:566–576CrossRefGoogle Scholar
  17. Djurdjevic’ L, Mitrović M, Pavlović P, Perišić S, Mačukanović-Jocić M (2005) Total phenolics and phenolic acids content in low (Chrysopogon gryllus) and mediocre quality (Festuca vallesiaca) forage grasses of Deliblato Sands meadow-pasture communities in Serbia. Czech J Anim Sci 50:54–59CrossRefGoogle Scholar
  18. Ergȕl FE, Sargın S, Ȍngen G, Sukan FV (2009) Dephenolisation of olive mill wastewater using adapted Trametes versicolor. Int Biodeterior Biodegrad 63:1–6CrossRefGoogle Scholar
  19. Fletcher JS, Hegde RS (1995) Release of phenols by perennial plant roots and their potential importance in bioremediation. Chemosphere 31:3009–3016CrossRefGoogle Scholar
  20. Gerhardt KE, Huang X-D, Glick BR, Greenberg BM (2009) Phytoremediation and rhizoremediation of organic soil contaminants: potential and challenges. Plant Sci 176:20–30CrossRefGoogle Scholar
  21. Gerhardt KE, Gerwing PD, Greenberg BM (2017) Opinion: taking phytoremediation from proven technology to accepted practice. Plant Sci 256:170–185CrossRefGoogle Scholar
  22. Gopalakrishnan S, Subbarao GV, Nakahara K, Yoshihashi T, Ito O, Maeda I, Ono H, Yoshida M (2007) Nitrification inhibitors from the root tissues of Brachiaria humidicola, a tropical grass. J Agric Food Chem 55:1385–1388CrossRefGoogle Scholar
  23. Harvey PJ, Campanella BF, Castro PM, Harms H, Lichtfouse E, Schäffner AR, Smrcek S, Werck-Reichhart D (2002) Phytoremediation of polyaromatic hydrocarbons, anilines and phenols. Environ Sci Pollut Res 9:29–47CrossRefGoogle Scholar
  24. Ibañéz SG, Wevar Oller AL, Paisio CE, Sosa Alderete LG, González PS, Medina MI, Agostini E (2017) The challenges of remediating metals using phytotechnologies. In: Donatti ER (ed) Heavy metals in the environment: microorganisms and bioremediation. CRC Press, Taylor & Francis, pp 173–191Google Scholar
  25. Juhanson J, Truu J, Heinaru E, Heinaru A (2009) Survival and catabolic performance of introduced Pseudomonas strains during phytoremediation and bioaugmentation field experiment. FEMS Microbiol Ecol 70:446–455CrossRefGoogle Scholar
  26. Karpouzas DG, Rousidou C, Papadopoulou KK, Bekris F, Zervakis GI, Singh BK, Ehaliotis C (2009) Effect of continuous olive mill wastewater applications in the presence and absence of nitrogen fertilization on the structure of rhizosphere-soil fungal communities. FEMS Microbiol Ecol 70:388–401CrossRefGoogle Scholar
  27. Keen BA, Raczkowski H (1921) The relation between the clay content and certain physical properties of a soil. J Agric Sci 11:441–449CrossRefGoogle Scholar
  28. Kerley MS, Pahey GC, Gould JM, Iannotti EL (1988) Effects of lignification, cellulose crystallinity and enzyme accessible space on the digestibility of plant cell wall carbohydrates by the ruminant. Food Microstruct 7:59–65Google Scholar
  29. Khongkhaem P, Intasiri A, Luepromchai E (2011) Silica-immobilized Methylobacterium sp. NP3 and Acinetobacter sp. PK1 degrade high concentrations of phenol. J Appl Microbiol 52:448–455CrossRefGoogle Scholar
  30. Kietkwanboot A, Tran HT, Suttinun O (2015) Simultaneous dephenolization and decolorization of treated palm oil mill effluent by oil palm fiber-immobilized Trametes hirsuta strain AK 04. Water Air Soil Pollut 226(345):1–13Google Scholar
  31. Kim SJ, Lim JM, Hamada M, Ahn JH, Weon HY, Suzuki KI, Ahn TY, Kwon SW (2015) Marmoricola solisilvae sp. nov. and Marmoricola terrae sp. nov., isolated from soil and emended description of the genus Marmoricola. Int J Syst Evol Microbiol 65:1825–1830CrossRefGoogle Scholar
  32. Kumar PA, Srinivas TNR, Sasikala C, Ramana CV, Imhoff JF (2008) Thiophaeococcus mangrove gen. nov., sp. nov., a photosynthetic, marine gammaproteobacterium isolated from the Bhitarkanika mangrove forest of India. Int J Syst Evol Microbiol 58:2660–2664CrossRefGoogle Scholar
  33. Liu BB, Chen W, Chu X, Yang Y, Salam N, Hu WY, Gao R, Duan YQ, Li WJ (2016) Mariniluteicoccus endophyticus sp. nov., an endophytic actinobacterium isolated from root of Ocimum basilicum. Int J Syst Evol Microbiol 66:1306–1310CrossRefGoogle Scholar
  34. Mostafa FIY, Helling CS (2003) Isolation and 16S DNA characterization of soil microorganisms from tropical soils capable of utilizing the herbicides hexazinone and tebuthiuron. J Environ Sci Health B 38:783–797CrossRefGoogle Scholar
  35. Namiki S, Otani T, Motoki Y, Seike N, Iwafune T (2018) Differential uptake and translocation of organic chemicals by several plant species from soil. J Pestic Sci 43:96–107CrossRefGoogle Scholar
  36. Neilson JW, Jordan FL, Maier RM (2013) Analysis of artifacts suggests DGGE should not be used for quantitative diversity analysis. J Microbiol Methods 92:256–263CrossRefGoogle Scholar
  37. Neoh CH, Lam CY, Lim CK, Yahya A, Ibrahim Z (2014) Decolorization of palm oil mill effluent using growing cultures of Curvularia clavata. Environ Sci Pollut Res 21:4397–4408CrossRefGoogle Scholar
  38. Oswal N, Sarma PM, Zinjarde SS, Pant A (2002) Palm oil mill effluent treatment by a tropical marine yeast. Bioresour Technol 85:35–37CrossRefGoogle Scholar
  39. Oviasogie PO, Aghimien AE (2003) Macronutrient status and speciation of Cu, Fe, Zn and Pb in soil containing palm oil mill effluent. Global J Pure Appl Sci 9:71–80Google Scholar
  40. Phenrat T, Teeratitayangkul P, Prasertsung I, Parichatprecha R, Jitsangiam P, Chomchalow N, Wichai S (2017) Vetiver plantlets in aerated system degrade phenol in illegally dumped industrial wastewater by phytochemical and rhizomicrobial degradation. Environ Sci Pollut Res 24:13235–13246CrossRefGoogle Scholar
  41. Phonepaseuth P, Rakkiatsakul V, Kachenchart B, Suttinun O, Luepromchai E (2019) Phenolic compounds removal by grasses and soil bacteria after land application of treated palm oil mill effluent: a pot study. Environ Eng Res 24:127–136CrossRefGoogle Scholar
  42. Proestos C, Komaitis M (2008) Application of microwave-assisted extraction to the fast extraction of plant phenolic compounds. LWT 41:652–659CrossRefGoogle Scholar
  43. Raskin I, Ensley BD (2000) Phytoremediation of toxic metals: using plants to clean up the environment. Wiley-Interscience, New YorkGoogle Scholar
  44. Schmidt U (2003) Enhancing phytoextraction: the effect of chemical soil manipulation on mobility, plant accumulation, and leaching of heavy metals. J Environ Qual 32:1939–1954CrossRefGoogle Scholar
  45. Sheoran V, Sheoran A, Poonia P (2011) Role of hyperaccumulators in phytoextraction of metals from contaminated mining sites: a review. Crit Rev Environ Sci Technol 41:168–214CrossRefGoogle Scholar
  46. Silva KR, Salles JF, Seldin L, Elsas JD (2003) Application of a novel Paenibacillus-specific PCR-DGGE method and sequence analysis to assess the diversity of Paenibacillus spp. in the maize rhizosphere. J Microbiol Methods 54:213–231CrossRefGoogle Scholar
  47. Tang JC, Wang RG, Niu XW, Wang M, Chu HR, Zhou QX (2010) Characterisation of the rhizoremediation of petroleum-contaminated soil: effect of different influencing factors. Biogeosciences 7:3961–3969CrossRefGoogle Scholar
  48. Teng Y, Shen Y, Luo Y, Sun X, Sun M, Fu D, Li Z, Christie P (2011) Influence of Rhizobium meliloti on phytoremediation of polycyclic aromatic hydrocarbons by alfalfa in an aged contaminated soil. J Hazard Mater 186:1271–1276CrossRefGoogle Scholar
  49. Ulappa AC, Kelsey RG, Frye GG, Rachlow JL, Shipley LA, Bond L, Pu X, Forbey JS (2014) Plant protein and secondary metabolites influence diet selection in a mammalian specialist herbivore. J Mammal 95(4):834–842CrossRefGoogle Scholar
  50. Uteau D, Hafner S, Pagenkemper SK, Peth S, Wiesenberg GLB, Kuzyakov Y, Horn R (2015) Oxygen and redox potential gradients in the rhizosphere of alfalfa grown on a loamy soil. J Plant Nutr Soil Sci 178:278–287CrossRefGoogle Scholar
  51. Ventorino V, Sannino F, Piccolo A, Cafaro V, Carotenuto R, Pepe O (2014) Methylobacterium populi VP2: plant growth-promoting bacterium isolated from a highly polluted environment for polycyclic aromatic hydrocarbon (PAH) biodegradation. Sci World J 2014(931793):1–11CrossRefGoogle Scholar
  52. Vinatoru M, Toma M, Radu O, Filip PI, Lazurca D, Mason TJ (1997) The use of ultrasound for the extraction of bioactive principles from plant materials. Ultrason Sonochem 4:135–139CrossRefGoogle Scholar
  53. Wrenn BA, Venosa AD (1996) Selective enumeration of aromatic and aliphatic hydrocarbon degrading bacteria by a most-probable number procedure. Can J Microbiol 42:252–258CrossRefGoogle Scholar
  54. Wu TY, Mohammad AW, Jahim JM, Anuar N (2009) A holistic approach to managing palm oil mill effluent (POME): biotechnological advances in the sustainable reuse of POME. Biotechnol Adv 27:40–52CrossRefGoogle Scholar
  55. Wu XY, Xu XM, Fu SY, Yuan JP, Peng J, Yan TP, Wu CF, Wang JH (2012) Impact of different polymerase chain reaction (PCR) strategies on denaturing gradient gel electrophoresis-based analysis of bacterial communities in soils/sediments from the Northern Jiangsu Oil Field, China. Afr J Microbiol Res 6:7094–7102Google Scholar
  56. Zahan KA, Kano M (2018) Biodiesel production from palm oil, its by-products, and mill effluent: a review. Energies 11(2132):1–25Google Scholar
  57. Zhivotovsky OP, Kuzovkina YA, Schulthess CP, Morris T, Pettinelli D (2011) Lead uptake and translocation by willows in pot and field experiment. Int J Phytoremediat 13:731–749CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.Environmental Assessment and Technology for Hazardous Waste Management Research Center, Faculty of Environmental ManagementPrince of Songkla UniversitySongkhlaThailand
  2. 2.Department of Plant Science, Faculty of Natural ResourcesPrince of Songkla UniversitySongkhlaThailand
  3. 3.Microbial Technology for Marine Pollution Treatment Research Unit, Department of Microbiology, Faculty of ScienceChulalongkorn UniversityBangkokThailand
  4. 4.Center of Excellence on Hazardous Substance Management (HSM)BangkokThailand

Personalised recommendations