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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, 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. To investigate, 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 subjectedmore » 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.« less

Authors:
 [1];  [2];  [2];  [2];  [2]
  1. Colorado State Univ., Pueblo, CO (United States)
  2. Univ. of Arizona, Tucson, AZ (United States)
Publication Date:
Research Org.:
Univ. of Arizona, Tucson, AZ (United States)
Sponsoring Org.:
USDOE Office of Energy Efficiency and Renewable Energy (EERE), Sustainable Transportation Office. Bioenergy Technologies Office
OSTI Identifier:
1592755
Grant/Contract Number:  
EE0006269; EE0003046
Resource Type:
Accepted Manuscript
Journal Name:
Journal of Applied Phycology
Additional Journal Information:
Journal Volume: 31; Journal Issue: 2; Journal ID: ISSN 0921-8971
Publisher:
Springer
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Chlorophyta; Cyanobacteria; DOE1412; Molecular detection; Polymerase chain reaction

Citation Formats

Park, Sang-Hyuck, Steichen, Seth A., Li, Xuehui, Ogden, Kimberly, and Brown, Judith K. Association of Vampirovibrio chlorellavorus with decline and death of Chlorella sorokiniana in outdoor reactors. United States: N. p., 2018. Web. doi:10.1007/s10811-018-1633-9.
Park, Sang-Hyuck, Steichen, Seth A., Li, Xuehui, Ogden, Kimberly, & Brown, Judith K. Association of Vampirovibrio chlorellavorus with decline and death of Chlorella sorokiniana in outdoor reactors. United States. https://doi.org/10.1007/s10811-018-1633-9
Park, Sang-Hyuck, Steichen, Seth A., Li, Xuehui, Ogden, Kimberly, and Brown, Judith K. Mon . "Association of Vampirovibrio chlorellavorus with decline and death of Chlorella sorokiniana in outdoor reactors". United States. https://doi.org/10.1007/s10811-018-1633-9. https://www.osti.gov/servlets/purl/1592755.
@article{osti_1592755,
title = {Association of Vampirovibrio chlorellavorus with decline and death of Chlorella sorokiniana in outdoor reactors},
author = {Park, Sang-Hyuck and Steichen, Seth A. and Li, Xuehui and Ogden, Kimberly and Brown, Judith K.},
abstractNote = {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, 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. To investigate, 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.},
doi = {10.1007/s10811-018-1633-9},
journal = {Journal of Applied Phycology},
number = 2,
volume = 31,
place = {United States},
year = {Mon Oct 01 00:00:00 EDT 2018},
month = {Mon Oct 01 00:00:00 EDT 2018}
}

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Works referencing / citing this record:

Real-time quantitative detection of Vampirovibrio chlorellavorus, an obligate bacterial pathogen of Chlorella sorokiniana
journal, October 2018


Vampirovibrio chlorellavorus draft genome sequence, annotation, and preliminary characterization of pathogenicity determinants
journal, July 2019

  • Hovde, Blake T.; Steichen, Seth A.; Starkenburg, Shawn R.
  • Phycological Research, Vol. 68, Issue 1
  • DOI: 10.1111/pre.12392