Treatment of recalcitrant wastewater from ethanol and citric acid production using the microalga Chlorella vulgaris and the macrophyte Lemna minuscula

Water Res. 2002 Oct;36(17):4185-92. doi: 10.1016/s0043-1354(02)00143-4.


Laboratory-scale experiments were performed to develop a procedure for biological treatment of recalcitrant anaerobic industrial effluent (from ethanol and citric acid production) using first the microalga Chlorella vulgaris followed by the macrophyte Lemna minuscula. This recalcitrant dark-colored wastewater, containing high levels of organic matter and low pH, prevents the growth of microalgae and macrophytes, and therefore, could not be treated by them. Therefore, the wastewater was diluted to 10% of the original concentration with wash water from the production line. Within 4 days of incubation in the wastewater, C. vulgaris population grew from 5 x 10(5) to 2 x 10(6) cells/mL. This culture reduced ammonium ion (71.6%), phosphorus (28%), and chemical oxygen demand (COD) (61%), and dissolved a floating microbial biofilm after 5 days of incubation. Consequently, L. minuscule was able to grow in the treated wastewater (from 7 to 14 g/bioreactor after 6 days), precipitated the microalgal cells (by shading the culture), and reduced other organic matter and color (up to 52%) after an additional 6 days of incubation. However, L. minuscula did not improve removal of nutrients. This study demonstrates the feasibility of combining microalgae and macrophytes for bioremediation of recalcitrant industrial wastewater.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anaerobiosis
  • Araceae / metabolism*
  • Biodegradation, Environmental
  • Biofilms
  • Bioreactors*
  • Chlorella / metabolism*
  • Citric Acid*
  • Ethanol*
  • Phosphorus / metabolism
  • Quaternary Ammonium Compounds / metabolism
  • Time Factors
  • Water Microbiology
  • Water Purification / methods*


  • Quaternary Ammonium Compounds
  • Phosphorus
  • Citric Acid
  • Ethanol