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Journal of Applied Phycology

, Volume 31, Issue 2, pp 1131–1142 | Cite as

Association of Vampirovibrio chlorellavorus with decline and death of Chlorella sorokiniana in outdoor reactors

  • Sang-Hyuck Park
  • Seth A. Steichen
  • Xuehui Li
  • Kimberly Ogden
  • Judith K. BrownEmail author
Article

Abstract

The outdoor ARID raceway was established for optimizing the cultivation of microalgae for biofuel production. During the summers of 2014 and 2015, discoloration was observed in cultures of Chlorella sorokiniana (DOE1412), which shifted from a vibrant green color to yellow, followed by cell clumping, decline in density, and rapid death, resulting in 40–60% reduced biomass production. Total DNA was purified from the raceway samples and subjected to polymerase chain reaction (PCR) amplification using degenerate primers that amplify the 16S rRNA gene of eubacteria. BLASTn analysis of the cloned amplicon sequences revealed the presence of the Gram-negative, predatory bacterium, Vampirovibrio chlorellavorus. Scanning electron microscopic examination showed an abundance of coccoid cells, 0.3–0.6 μm in diameter, some of which were attached to C. sorokiniana cells. PCR amplification indicated the presence of V. chlorellavorus in raceway vessels, water lines, connective tubing, and in early, scaled-up DOE1412 cultures used to inoculate the raceway. Based on PCR detection, the decontamination of the equipment and water line with “Wal-Clean” more effectively eliminated V. chlorellavorus and delayed the onset of attack, compared to the chlorine disinfectant, trichloromelamine (TCM). Total DNA was isolated from soil samples collected monthly from the nearby Rillito River during 2014–2015 and subjected to PCR amplification using primers designed to amplify the 16S rRNA and 18S rRNA gene of V. chlorellavorus and C. sorokiniana, respectively. Results indicated that V. chlorellavorus and Chlorella spp. were present in most of the riverbed samples nearly year round, suggesting a possible naturally occurring reservoir of the predatory bacterium.

Keywords

Chlorophyta Cyanobacteria DOE1412 Molecular detection Polymerase chain reaction 

Notes

Acknowledgments

The authors thank Dr. Juergen Polle at the City University of New York, Brooklyn for providing the DOE1412 culture. We also thank the UA-ARID reactor cultivation and biology team members who provided outstanding support and enabled this research to be carried out.

Funding information

This project was funded by the USDOE Office of Biomass Program, DE-EE0003046, as part of the National Alliance for Advanced Biofuels and Bioproducts or NAABB consortium, and recently, by the DOE RAFT Project DE-EE0006269.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10811_2018_1633_MOESM1_ESM.docx (87 kb)
ESM 1 (DOCX 86 kb)

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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Sang-Hyuck Park
    • 1
  • Seth A. Steichen
    • 2
  • Xuehui Li
    • 3
  • Kimberly Ogden
    • 3
  • Judith K. Brown
    • 2
    Email author
  1. 1.Department of BiologyColorado State UniversityPuebloUSA
  2. 2.School of Plant SciencesThe University of ArizonaTucsonUSA
  3. 3.Department of Chemical and Environmental EngineeringThe University of ArizonaTucsonUSA

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