Abstract

The presence of heterotrophic bacteria in microalgal cultures can dilute the microalgal content of the harvested biomass, compete for nutrients, and be associated with culture crashes. Being able to detect and quantify heterotrophic bacteria would be of high value for monitoring culture health and reducing deleterious effects. Here, we developed and applied a new method that combines flow cytometry (FC) and fluorescence activated cell sorting (FACS) for the quantification of heterotrophic bacteria in cultures of the cyanobacterium Synechocystis sp. PCC 6803. Particles not containing chlorophyll – heterotrophic bacteria and cell debris – were separated from mixed cultures using FACS based on autofluorescence of Synechocystis. Heterotrophic bacteria were differentiated from cell debris using FC with SYTOX green fluorescence. Using microscopy, we verified that FACS was able to quantify heterotrophic bacteria in Synechocystis cultures effectively. Applying these methods to batch cultures of Synechocystis showed that the count proportions of heterotrophic bacteria were significant (3–13%) and that depletion of inorganic P in the culture favored Synechocystis over heterotrophic bacteria, but led to more cell lysis.

Original languageEnglish (US)
Pages (from-to)94-100
Number of pages7
JournalAlgal Research
Volume30
DOIs
StatePublished - Mar 1 2018

Fingerprint

Synechocystis sp. PCC 6803
flow cytometry
microscopy
Synechocystis
bacteria
mixed culture
cells
Cyanobacteria
fluorescence
chlorophyll
monitoring
biomass
nutrients
methodology

Keywords

  • Cyanobacteria
  • Flow cytometry
  • Heterotrophic bacteria
  • Quantification
  • Synechocystis sp. PCC 6803

ASJC Scopus subject areas

  • Agronomy and Crop Science

Cite this

Quantification of heterotrophic bacteria during the growth of Synechocystis sp. PCC 6803 using fluorescence activated cell sorting and microscopy. / Zhou, Yun; Eustance, Everett; Straka, Levi; Lai, Yen Jung Sean; Xia, Siqing; Rittmann, Bruce.

In: Algal Research, Vol. 30, 01.03.2018, p. 94-100.

Research output: Contribution to journalArticle

Zhou, Yun ; Eustance, Everett ; Straka, Levi ; Lai, Yen Jung Sean ; Xia, Siqing ; Rittmann, Bruce. / Quantification of heterotrophic bacteria during the growth of Synechocystis sp. PCC 6803 using fluorescence activated cell sorting and microscopy. In: Algal Research. 2018 ; Vol. 30. pp. 94-100.
@article{ddf3e71da83b4b679a3a31fa72b1d01d,
title = "Quantification of heterotrophic bacteria during the growth of Synechocystis sp. PCC 6803 using fluorescence activated cell sorting and microscopy",
abstract = "The presence of heterotrophic bacteria in microalgal cultures can dilute the microalgal content of the harvested biomass, compete for nutrients, and be associated with culture crashes. Being able to detect and quantify heterotrophic bacteria would be of high value for monitoring culture health and reducing deleterious effects. Here, we developed and applied a new method that combines flow cytometry (FC) and fluorescence activated cell sorting (FACS) for the quantification of heterotrophic bacteria in cultures of the cyanobacterium Synechocystis sp. PCC 6803. Particles not containing chlorophyll – heterotrophic bacteria and cell debris – were separated from mixed cultures using FACS based on autofluorescence of Synechocystis. Heterotrophic bacteria were differentiated from cell debris using FC with SYTOX green fluorescence. Using microscopy, we verified that FACS was able to quantify heterotrophic bacteria in Synechocystis cultures effectively. Applying these methods to batch cultures of Synechocystis showed that the count proportions of heterotrophic bacteria were significant (3–13{\%}) and that depletion of inorganic P in the culture favored Synechocystis over heterotrophic bacteria, but led to more cell lysis.",
keywords = "Cyanobacteria, Flow cytometry, Heterotrophic bacteria, Quantification, Synechocystis sp. PCC 6803",
author = "Yun Zhou and Everett Eustance and Levi Straka and Lai, {Yen Jung Sean} and Siqing Xia and Bruce Rittmann",
year = "2018",
month = "3",
day = "1",
doi = "10.1016/j.algal.2018.01.006",
language = "English (US)",
volume = "30",
pages = "94--100",
journal = "Algal Research",
issn = "2211-9264",
publisher = "Elsevier BV",

}

TY - JOUR

T1 - Quantification of heterotrophic bacteria during the growth of Synechocystis sp. PCC 6803 using fluorescence activated cell sorting and microscopy

AU - Zhou, Yun

AU - Eustance, Everett

AU - Straka, Levi

AU - Lai, Yen Jung Sean

AU - Xia, Siqing

AU - Rittmann, Bruce

PY - 2018/3/1

Y1 - 2018/3/1

N2 - The presence of heterotrophic bacteria in microalgal cultures can dilute the microalgal content of the harvested biomass, compete for nutrients, and be associated with culture crashes. Being able to detect and quantify heterotrophic bacteria would be of high value for monitoring culture health and reducing deleterious effects. Here, we developed and applied a new method that combines flow cytometry (FC) and fluorescence activated cell sorting (FACS) for the quantification of heterotrophic bacteria in cultures of the cyanobacterium Synechocystis sp. PCC 6803. Particles not containing chlorophyll – heterotrophic bacteria and cell debris – were separated from mixed cultures using FACS based on autofluorescence of Synechocystis. Heterotrophic bacteria were differentiated from cell debris using FC with SYTOX green fluorescence. Using microscopy, we verified that FACS was able to quantify heterotrophic bacteria in Synechocystis cultures effectively. Applying these methods to batch cultures of Synechocystis showed that the count proportions of heterotrophic bacteria were significant (3–13%) and that depletion of inorganic P in the culture favored Synechocystis over heterotrophic bacteria, but led to more cell lysis.

AB - The presence of heterotrophic bacteria in microalgal cultures can dilute the microalgal content of the harvested biomass, compete for nutrients, and be associated with culture crashes. Being able to detect and quantify heterotrophic bacteria would be of high value for monitoring culture health and reducing deleterious effects. Here, we developed and applied a new method that combines flow cytometry (FC) and fluorescence activated cell sorting (FACS) for the quantification of heterotrophic bacteria in cultures of the cyanobacterium Synechocystis sp. PCC 6803. Particles not containing chlorophyll – heterotrophic bacteria and cell debris – were separated from mixed cultures using FACS based on autofluorescence of Synechocystis. Heterotrophic bacteria were differentiated from cell debris using FC with SYTOX green fluorescence. Using microscopy, we verified that FACS was able to quantify heterotrophic bacteria in Synechocystis cultures effectively. Applying these methods to batch cultures of Synechocystis showed that the count proportions of heterotrophic bacteria were significant (3–13%) and that depletion of inorganic P in the culture favored Synechocystis over heterotrophic bacteria, but led to more cell lysis.

KW - Cyanobacteria

KW - Flow cytometry

KW - Heterotrophic bacteria

KW - Quantification

KW - Synechocystis sp. PCC 6803

UR - http://www.scopus.com/inward/record.url?scp=85042927478&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85042927478&partnerID=8YFLogxK

U2 - 10.1016/j.algal.2018.01.006

DO - 10.1016/j.algal.2018.01.006

M3 - Article

AN - SCOPUS:85042927478

VL - 30

SP - 94

EP - 100

JO - Algal Research

JF - Algal Research

SN - 2211-9264

ER -