Microalgae growth-promoting bacteria as “helpers” for microalgae: A novel approach for removing ammonium and phosphorus from municipal wastewater

  • L. E. de-Bashan
  • J. P. Hernandez
  • Y. Bashan
Part of the Developments in Plant and Soil Sciences book series (DPSS, volume 102)


A combination of microalgae (Chlorella vulgaris or C. sorokiniana) and a microalgae growth-promoting bacterium (MGPB, Azospirillum brasilense strain Cd), co-immobilized in small alginate beads, was developed to remove nutrients (P and N) from municipal wastewater. This paper describes the most recent technical details necessary for successful co-immobilization of the two microorganisms, and the usefulness of the approach in cleaning the municipal wastewater of the city of La Paz, Mexico. A. brasilense Cd significantly enhanced the growth of both Chlorella species when the co-immobilized microorganisms were grown in wastewater. A. brasilense is incapable of significant removal of nutrients from the wastewater, whereas both microalgae can. Co-immobilization of the two microorganisms was superior to removal by the microalgae alone, reaching removal of up to 100% ammonium, 94% nitrate, and 92% phosphorus within 6 days (varied with the source of the wastewater), compared to 75% ammonium, 84% nitrate, and 89% phosphorus by the microalgae alone. This study shows the potential of co-immobilization of microorganisms in small beads to serve as a treatment for wastewater.

Key words

phosphate nitrate microalgae growth promoting bacteria wastewater 


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  1. APHA, AWWA, WPCF (American Public Health Association, American Waterworks Association. Water Pollution Control Federation) 1992 Standard methods for the examination of water and wastewater 17th ed. Madrid, Spain. 1105 p.Google Scholar
  2. Bashan Y 1986 Alginate beads as synthetic inoculant carriers for the slow release of bacteria that affect plant growth. Appl. Environ. Microbiol. 51, 1089–1098.PubMedGoogle Scholar
  3. Bashan Y 1998 Inoculants of plant growth-promoting bacteria for use in agriculture. Biotechnol. Adv. 16, 729–770.CrossRefGoogle Scholar
  4. Bashan Y, Hernandez J-P, Leyva L A and Bacilio M 2002 Alginate microbeads as inoculant carrier for plant growthpromoting bacteria. Biol. Fertil. Soils 35, 359–368.CrossRefGoogle Scholar
  5. Bashan Y and Holguin G 1997 Azospirillum-plant relationships: environmental and physiological advances (1990–1996). Can. J. Microbiol. 43, 103–121.CrossRefGoogle Scholar
  6. Bashan Y, Holguin G and Lifshitz R 1993 Isolation and characterization of plant growth-promoting rhizobacteria. In Methods in Plant Molecular Biology and Biotechnology. Eds. B R Glick and J E Thompson. pp. 331–345. CRC Press, Boca Raton, FL.Google Scholar
  7. Chevalier P and De la Noüe J 1985 Wastewater nutrient removal with microalgae immobilized in carrageenan. Enzyme Microbiol. Technol. 7, 621–624.CrossRefGoogle Scholar
  8. de-Bashan L E, Bashan Y, Moreno M, Lebsky V K and Bustillos J J 2002a Increased pigment and lipid content, lipid variety, and cell and population size of the microalgae Chlorella spp. when co-immobilized in alginate beads with the microalgae-growth-promoting bacterium Azospirillum brasilense. Can. J. Microbiol. 48, 514–521.PubMedCrossRefGoogle Scholar
  9. de-Bashan L E, Moreno M, Hernandez J-P and Bashan Y 2002b Ammonium and phosphorus removal from continuous and semi-continuous cultures by the microalgae Chlorella vulgaris coimmobilized in alginate beads with Azospirillum brasilense. Water Res. 36, 2941–2948.PubMedCrossRefGoogle Scholar
  10. De la Noüe J and De Pauw N 1988 The potential of microalgal biotechnology: a review of production and uses of microalgae. Biotechnol. Adv. 6, 725–770.PubMedCrossRefGoogle Scholar
  11. Gonzalez L E and Bashan Y 2000 Growth promotion of the microalgae Chlorella vulgaris when coimmobilized and cocultured in alginate beads with the plant growth-promoting bacteria Azospirillum brasilense. Appl. Environ. Microbiol. 66, 1537–1541.Google Scholar
  12. Gonzalez L E, Cañizares R O and Baena S 1997 Efficiency of ammonia and phosphorus removal from a Colombian agroindustrial wastewater by the microalgae Chlorella vulgaris and Scenedesmus dimorphus. Biores. Technol. 60, 259–262.CrossRefGoogle Scholar
  13. Gonzalez-Bashan L E, Lebsky V, Hernandez J P, Bustillos J J and Bashan Y 2000 Changes in the metabolism of the microalgae Chlorella vulgaris when coimmobilized in alginate with the nitrogen-fixing Phyllobacterium myrsinacearum. Can. J. Microbiol. 46, 653–659.PubMedCrossRefGoogle Scholar
  14. Lau P, Tam N F Y and Wong Y S 1997 Wastewater nutrients (N and P) removal by carrageenan and alginate immobilized Chlorella vulgaris. Environ. Technol. 18, 893–899.CrossRefGoogle Scholar
  15. Lebsky V K, Gonzalez-Bashan L E and Bashan Y 2001 Ultrastructure of coimmobilization of the microalga Chlorella vulgaris with the plant growth-promoting bacterium Azospirillum brasilense and with its natural associative bacterium Phyllobacterium myrsinacearum in alginate beads. Can. J. Microbiol. 47, 1–8.PubMedCrossRefGoogle Scholar
  16. Mouget J L, Dakhama A and De Lavoie la Noüe M C J 1995 Algal growth enhancement by bacteria: is consumption of photosynthetic oxygen involved? FEMS Microbiol. Ecol. 18, 35–44.CrossRefGoogle Scholar
  17. Ogbonna J C, Yoshizawa H and Tanaka H 2000 Treatment of high strength organic wastewater by a mixed culture of photosynthetic microorganisms. J. Appl. Phycol. 12, 277–284.CrossRefGoogle Scholar
  18. Tam N F Y and Wong Y S 2000 Effect of immobilized microalgal bead concentration on wastewater nutrient removal. Environ. Pollut. 107, 145–151.PubMedCrossRefGoogle Scholar
  19. Valderrama L T, Del Campo C M, Rodriguez C M, de-Bashan L E and Bashan Y 2002 Treatment of recalcitrant wastewater from ethanol and citric acid production using the microalga Chlorella vulgaris and the macrophyte Lemna minuscula. Water Res. 36, 4185–4192.PubMedCrossRefGoogle Scholar

Copyright information

© Springer 2007

Authors and Affiliations

  • L. E. de-Bashan
    • 1
  • J. P. Hernandez
    • 1
  • Y. Bashan
    • 1
  1. 1.The Center for Biological Research of the Northwest (CIB)The Environmental Microbiology GroupLa PazMexico

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