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

Sang Hyuck Park, Seth A. Steichen, Xuehui Li, Kimberly L Ogden, Judith K Brown

Research output: Contribution to journalArticle

4 Citations (Scopus)

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.

Original languageEnglish (US)
JournalJournal of Applied Phycology
DOIs
StateAccepted/In press - Jan 1 2018

Fingerprint

Chlorella sorokiniana
raceways
polymerase chain reaction
death
amplification
ribosomal RNA
shoreline
DNA
bacterium
gene
Eubacteria
biofuel
bacteria
disinfectants
Chlorella
decontamination
chlorine
cells
stream channels
discoloration

Keywords

  • Chlorophyta
  • Cyanobacteria
  • DOE1412
  • Molecular detection
  • Polymerase chain reaction

ASJC Scopus subject areas

  • Aquatic Science
  • Plant Science

Cite this

Association of Vampirovibrio chlorellavorus with decline and death of Chlorella sorokiniana in outdoor reactors. / Park, Sang Hyuck; Steichen, Seth A.; Li, Xuehui; Ogden, Kimberly L; Brown, Judith K.

In: Journal of Applied Phycology, 01.01.2018.

Research output: Contribution to journalArticle

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title = "Association of Vampirovibrio chlorellavorus with decline and death of Chlorella sorokiniana in outdoor reactors",
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.",
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