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Carboxymethyl cellulose and Pluronic F68 protect the dinoflagellate Protoceratium reticulatum against shear-associated damage

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Abstract

The red-tide dinoflagellate Protoceratium reticulatum is shown to be protected against turbulence-associated damage by the use of the additives Pluronic F68 (PF68) and carboxymethyl cellulose (CMC) in the culture medium. Relative to agitated controls, these additives had a dose-dependent protective effect at concentrations of up to 0.4 and 0.5 g L−1 for CMC and F68, respectively. In static cultures, these additives inhibited growth directly or indirectly at a concentration of >0.5 g L−1. Compared to CMC, PF68 was a better protectant overall. Cell-specific production of yessotoxins was enhanced under elevated shear stress regimens so long as the turbulence intensity was insufficient to damage the cells outright. Shear-induced production of reactive oxygen species and direct effects of turbulence on the cell cycle contributed to the observed shear effects.

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Abbreviations

CMC:

Carboxymethyl cellulose

d p :

Diameter of the cells (m)

d s :

Diameter of the shake flask (m)

n :

Rotational speed of the shake flask (s−1)

PF68:

Pluronic F68

ROS:

Reactive oxygen species

t c :

Total cycle time (s)

t s :

Duration of the quiescent period within one agitation cycle (s)

t t :

Duration of turbulence within one agitation cycle (s)

V L :

Volume of the broth in the flask (m3)

YTX:

Yessotoxin

γ ar :

Reference value of average shear rate calculated using d p = 20.5 μm and seawater viscosity (s−1)

γ av :

Average shear rate (s−1)

γ t :

Applied shear rate (s−1)

μ L :

Viscosity of the broth (Pa s)

μ r :

Ratio of the maximum specific growth rates of the agitated culture and the corresponding static control

μ ra :

Ratio of the maximum specific growth rate in an agitated culture with protective additive and the maximum specific growth rate in an identically agitated culture but without the additive

ν :

Cycle frequency (= 1/t c) (s−1)

ρ L :

Density of the broth (kg m−3)

τ ar :

Reference value of average shear stress calculated using d p = 20.5 μm (Pa) and seawater viscosity (s−1)

τ av :

Average shear stress (Pa)

τ t :

Applied shear stress (Pa)

ϕ :

Fraction of the time shaken in one agitation cycle

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Acknowledgments

This research was supported by the Spanish Ministry of Education and Science (AGL2005-07924-C04-04) and the Spanish Ministry of Science and Innovation (CTQ2008-06754-C04-02/PPQ).

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Authors and Affiliations

  1. Department of Chemical Engineering, University of Almería, 04120, Almería, Spain

    J. J. Gallardo Rodríguez, A. Sánchez Mirón, F. García Camacho, M. C. Cerón García, E. H. Belarbi & E. Molina Grima

  2. School of Engineering, Massey University, Private Bag 11, 222, Palmerston North, New Zealand

    Y. Chisti

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  1. J. J. Gallardo Rodríguez
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  2. A. Sánchez Mirón
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  3. F. García Camacho
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  4. M. C. Cerón García
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  5. E. H. Belarbi
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  7. E. Molina Grima
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Correspondence to F. García Camacho.

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Gallardo Rodríguez, J.J., Sánchez Mirón, A., García Camacho, F. et al. Carboxymethyl cellulose and Pluronic F68 protect the dinoflagellate Protoceratium reticulatum against shear-associated damage. Bioprocess Biosyst Eng 34, 3–12 (2011). https://doi.org/10.1007/s00449-010-0441-7

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