Post Cryopreservation Growth Kinetic and Photosynthetic Assessment of an Acid Tolerant Strain of <i>Stichococcus bacillaris</i> — Oak Academic Publishing
Research ArticleOpen AccessGoogle Scholar indexed
Post Cryopreservation Growth Kinetic and Photosynthetic Assessment of an Acid Tolerant Strain of <i>Stichococcus bacillaris</i>
Department of Biology, University of Naples “Federico II”, Naples, Italy
,
Department of Chemical, Materials and Industrial Production Engineering, University of Naples “Federico II”, P.le V. Tecchio, Naples, Italy
,
Department of Biology, University of Naples “Federico II”, Naples, Italy
,
Department of Chemical, Materials and Industrial Production Engineering, University of Naples “Federico II”, P.le V. Tecchio, Naples, Italy
,
Department of Biology, University of Naples “Federico II”, Naples, Italy
,
Department of Biology, University of Naples “Federico II”, Naples, Italy
,
Department of Biology, University of Naples “Federico II”, Naples, Italy
,
Department of Biology, University of Naples “Federico II”, Naples, Italy
,
ACUF, Algal Collection of University of Naples “Federico II”, Naples, Italy
,
Institute for Marine Biological Resources and Biotechnologies, Italian National Research Council, CNR-IRBIM, Ancona, Italy
,
Earth-Life Science Institute, ELSI, Tokyo Institute of Technology, Tokyo, Japan
,
Marine Chemistry and Geochemistry Department, Woods Hole Oceanographic Institution, Woods Hole, MA, USA
,
Department of Marine and Coastal Science, Rutgers University, New Brunswick, NJ, USA
1 Department of Biology, University of Naples “Federico II”, Naples, Italy
2 Department of Chemical, Materials and Industrial Production Engineering, University of Naples “Federico II”, P.le V. Tecchio, Naples, Italy
3 Department of Biology, University of Naples “Federico II”, Naples, Italy
4 Department of Chemical, Materials and Industrial Production Engineering, University of Naples “Federico II”, P.le V. Tecchio, Naples, Italy
5 Department of Biology, University of Naples “Federico II”, Naples, Italy
6 Department of Biology, University of Naples “Federico II”, Naples, Italy
7 Department of Biology, University of Naples “Federico II”, Naples, Italy
8 Department of Biology, University of Naples “Federico II”, Naples, Italy
9 ACUF, Algal Collection of University of Naples “Federico II”, Naples, Italy
10 Institute for Marine Biological Resources and Biotechnologies, Italian National Research Council, CNR-IRBIM, Ancona, Italy
11 Earth-Life Science Institute, ELSI, Tokyo Institute of Technology, Tokyo, Japan
12 Marine Chemistry and Geochemistry Department, Woods Hole Oceanographic Institution, Woods Hole, MA, USA
13 Department of Marine and Coastal Science, Rutgers University, New Brunswick, NJ, USA
Preserving microbial diversity has become a strategic undertaking. Thus, <i>ex situ</i> microalgal culture conservation results in strategic and functional resource in both biodiversity protection and application domains. Cryopreservation of microalgae has been practiced since the 1960s and is now considered the optimal preservation strategy. Furthermore, the overall monitoring during growth of cultures after freezing/thawing protocols was hardly investigated and there is poor evaluation related to preserve especially the photosystem apparatus. The present study focuses on <i>Stichococcus bacillaris</i> as case study for short-term cryopreservation at −80 °C storage. Various freezing pretreatments using cryoprotective agents, and two thawing methods were compared introducing a novel variable to evaluate viability recovery and assessing growth kinetics of cultures immediately after thawing and after a series batch cultivation. Photosynthetic rate and pigments assessment were proposed to evaluate hidden metabolic cell damage. Results underline cryoprotective agents can increase the kinetic recovery of preserved cells in terms of reduction of lag phase during batch cultivation tests: the use of dimethyl sulfoxide and glycerol granted a growth comparable to unpreserved cells when sudden thawing occurs after 24 hours of storage, but recovery after preservation is less sensitive to cryoprotective agents when gradual thawing and 1 month of storage is considered. However, cells are always able to restore their physiological pathways even without agents, so their kinetic effect has been proved and quantified. Interestingly, both the photosynthetic efficiency and the ratio between total chlorophyll and carotenoids are comparable (0.75 F<sub>v</sub>/F<sub>m</sub>, 2.2 ± 0.25 g/g) to unpreserved cells and they are unsensitive to chosen agents, but the ratio between chlorophyll <i>a</i> and chlorophyll <i>b </i>was clearly altered (up to 10 times), suggesting that photoactive pigments relative proportions can result in similar growth kinetic performances. Long-term studies will be carried out to assess whether the differences found could cause chronic damage to photosystem efficiency of <i>S. bacillaris</i> cultures.
Andersen, R.A. (2013) The Microalgal Cell. In: Richmond, A., Emeritus and Hu, Q., Eds., Handbook of Microalgal Culture : Applied Phycology and Biotechnology , 2nd Edition, Wiley, Hoboken, 3-20. https://doi.org/10.1002/9781118567166.ch1
Knoll, A.H., Javaux, E.J., Hewitt, D. and Cohen, P. (2006) Eukaryotic Organisms in Proterozoic Oceans. Philosophical Transactions of the Royal Society B : Biological Sciences , 361, 1023-1038. https://doi.org/10.1098/rstb.2006.1843
Guiry, M.D. (2024) How Many Species of Algae Are There? A Reprise. Four Kingdoms, 14 Phyla, 63 Classes and Still Growing. Journal of Phycology , 60, 214-228. https://doi.org/10.1111/jpy.13431
Masojídek, J., Torzillo, G. and Koblízek, M. (2013) Photosynthesis in Microalgae. In: Richmond, A., Emeritus and Hu, Q., Eds., Handbook of Microalgal Culture : A p plied Phycology and Biotechnology , 2nd Edition, Wiley, Hoboken, 21-36. https://doi.org/10.1002/9781118567166.ch2
Shimizu, Y. (1996) Microalgal Metabolites: A New Perspective. Annual Review of Microbiology , 50, 431-465. https://www.annualreviews.org https://doi.org/10.1146/annurev.micro.50.1.431
Mazhar, S., Cohen, J.D. and Hasnain, S. (2013) Auxin Producing Non-Heterocystous Cyanobacteria and Their Impact on the Growth and Endogenous Auxin Homeostasis of Wheat. Journal of Basic Microbiology , 53, 996-1003. https://doi.org/10.1002/jobm.201100563
Rangel-Yagui, C.D.O., Danesi, E.D.G., De Carvalho, J.C.M. and Sato, S. (2004) Chlorophyll Production from Spirulina platensis : Cultivation with Urea Addition by Fed-Batch Process. Bioresource Technology , 92, 133-141. https://doi.org/10.1016/j.biortech.2003.09.002
Soletto, D., Binaghi, L., Lodi, A., Carvalho, J.C.M. and Converti, A. (2005) Batch and Fed-Batch Cultivations of Spirulina platensis Using Ammonium Sulphate and Urea as Nitrogen Sources. Aquaculture , 243, 217-224. https://doi.org/10.1016/j.aquaculture.2004.10.005
Ponomarenko, L.P., et al . (2004) Sterols of Marine Microalgae Pyramimonas cf. cordata (Prasinophyta), Attheya ussurensis sp. nov. (Bacillariophyta) and a Spring Diatom Bloom from Lake Baikal. Comparative Biochemistry and Physiology Part B : Biochemistry and Molecular Biology , 138, 65-70. https://doi.org/10.1016/j.cbpc.2004.02.007
Luo, W., Du, W., Su, Y., Hui, J., Zhuang, J. and Liu, L. (2015) Growth Characteristic of the Oleaginous Mi-Croalga Chlorella ellipsoidea SD-0701 with Lipid Accumulation. Natural Resources , 6, 130-139. https://doi.org/10.4236/nr.2015.62012
Growth Kinetic
Photosynthetic Rate
Mutanda, T., Ramesh, D., Karthikeyan, S., Kumari, S., Anandraj, A. and Bux, F. (2011) Bioprospecting for Hyper-Lipid Producing Microalgal Strains for Sustainable Biofuel Production. Bioresource Technology , 102, 57-70. https://doi.org/10.1016/j.biortech.2010.06.077
Alvarez, A.L., Weyers, S.L., Goemann, H.M., Peyton, B.M. and Gardner, R.D. (2021) Microalgae, Soil and Plants: A Critical Review of Microalgae as Renewable Resources for Agriculture. Algal Research , 54, Article 102200. https://doi.org/10.1016/j.algal.2021.102200
Gaurav, N., Sivasankari, S., Kiran, G.S., Ninawe, A. and Selvin, J. (2017) Utilization of Bioresources for Sustainable Biofuels: A Review. Renewable and Sustainable Energy Reviews , 73, 205-214. https://doi.org/10.1016/j.rser.2017.01.070
Mingazzini, M. and Palumbo, M.T. (2015) Open Mass Cultures of Marine Microalgae for Biodiesel Production: Laboratory Approach to Study Species Competition in Mixed Cultures. Natural Resources , 6, 174-180. https://doi.org/10.4236/nr.2015.63016
Mata, T.M., Martins, A.A. and Caetano, N.S. (2010) Microalgae for Biodiesel Production and Other Applications: A Review. Renewable and Sustainable Energy R e views , 14, 217-232. https://doi.org/10.1016/j.rser.2009.07.020
Olivieri, G., Marzocchella, A., Andreozzi, R., Pinto, G. and Pollio, A. (2011) Biodiesel Production from Stichococcus Strains at Laboratory Scale. Journal of Chemical Technology and Biotechnology , 86, 776-783. https://doi.org/10.1002/jctb.2586
Prakash, O., Nimonkar, Y. and Shouche, Y.S. (2013) Practice and Prospects of Microbial Preservation. FEMS Microbiology Letters , 339, 1-9. https://doi.org/10.1111/1574-6968.12034
Lakeman, M.B., Von Dassow, P. and Cattolico, R.A. (2009) The Strain Concept in Phytoplankton Ecology. Harmful Algae , 8, 746-758. https://doi.org/10.1016/j.hal.2008.11.011
Hoefman, S., Van Hoorde, K., Boon, N., Vandamme, P., De Vos, P. and Heylen, K. (2012) Survival or Revival: Long-Term Preservation Induces a Reversible Viable But Non-Culturable State in Methane-Oxidizing Bacteria. PLOS ONE , 7, e34196. https://doi.org/10.1371/journal.pone.0034196
Foo, S.C., Mok, C.Y., Ho, S.Y. and Khong, N.M.H. (2023) Microalgal Culture Preservation: Progress, Trends and Future Developments. Algal Research , 71, Article 103007. https://doi.org/10.1016/j.algal.2023.103007
Day, J.G. and Fleck, R.A. (2015) Cryo-Injury in Algae and the Implications This Has to the Conservation of Micro-Algae. Microalgae Biotechnology , 1, 1-11. https://doi.org/10.1515/micbi-2015-0001
McGrath, M.S., Daggett, P.-M. and Dilworth, S. (1978) Freeze-Drying of Algae: Chlorophyta and Chrysophyta. Journal of Phycology , 14, 521-525. https://doi.org/10.1111/j.1529-8817.1978.tb02480.x
Kapoore, R.V., et al . (2019) Effects of Cryopreservation on Viability and Functional Stability of an Industrially Relevant Alga. Scientific Reports , 9, Article No. 2093. https://doi.org/10.1038/s41598-019-38588-6
Morschett, H., Reich, S., Wiechert, W. and Oldiges, M. (2016) Simplified Cryopreservation of the Microalga Chlorella Vulgaris Integrating a Novel Concept for Cell Viability Estimation. Engineering in Life Sciences , 16, 36-44. https://doi.org/10.1002/elsc.201500056
Kugler, A., Kumari, P., Kokabi, K., Itkin, M., Malitsky, S. and Khozin-Goldberg, I. (2020) Resilience to Freezing in the Vegetative Cells of the Microalga Lobosphaera incisa (Trebouxiophyceae, Chlorophyta). Journal of Phycology , 56, 334-345. https://doi.org/10.1111/jpy.12948
Garrido-Cardenas, J.A., Han, X., Alonso, D.L. and García-Maroto, F. (2019) Evaluation and Optimization of a Methodology for the Long-Term Cryogenic Storage of T e tradesmus obliquus at −80˚C. Applied Microbiology and Biotechnology , 103, 2381-2390. https://doi.org/10.1007/s00253-019-09650-0
Bui, T.V.L., Ross, I.L., Jakob, G. and Hankamer, B. (2013) Impact of Procedural Steps and Cryopreservation Agents in the Cryopreservation of Chlorophyte Microalgae. PLOS ONE , 8, e78668. https://doi.org/10.1371/journal.pone.0078668
Smith, D., Ryan, M.J. and Stackebrandt, E. (2008) The ex situ Conservation of Microorganisms: Aiming at a Certified Quality Management. In: Doelle, H.W. and DaSilva, E.J., Eds., Encyclopedia of Life Sciences Support , EoLSS, Oxford. https://www.researchgate.net/publication/266734069
D’Elia, L., Del Mondo, A., Santoro, M., De Natale, A., Pinto, G. and Pollio, A. (2018) Microorganisms from Harsh and Extreme Environments: A Collection of Living Strains at ACUF (Naples, Italy). Ecological Questions , 29, 63-74. https://doi.org/10.12775/EQ.2018.023
Rampelotto, P.H. (2013) Extremophiles and Extreme Environments. Life , 3, 482-485. https://www.mdpi.com/journal/life
De Luca, P., Taddei, R. and Varano, L. (1978) « Cyanidioschyzon Merolae »: A New Alga of Thermal Acidic Environments: « Cyanidioschyzon Merolae »: Una nuova alga di ambienti termali acidi. Webbia , 33, 37-44. https://doi.org/10.1080/00837792.1978.10670110
Pollio, A., Aliotta, G., Pinto, G., Paterno, M. and Bevilacqua, A. (1997) Ecophysiological Characters and Biochemical Composition of Stichococcus bacillaris NAEGELI Strains from Low pH Environments. Algological Studies / Archiv für Hydrobiologie , 84, 129-143. https://doi.org/10.1127/algol_stud/84/1997/129
Teoh, M.-L., Chu, W.-L., Marchant, H. and Phang, S.-M. (2004) Influence of Culture Temperature on the Growth, Biochemical Composition and Fatty Acid Profiles of Six Antarctic Microalgae. Journal of Applied Phycology , 16, 421-430. https://doi.org/10.1007/s10811-004-5502-3
Holm-Hansen, O. (1963) Viability of Blue-Green and Green Algae after Freezing. Physiologia Plantarum , 16, 530-540. https://doi.org/10.1111/j.1399-3054.1963.tb08330.x
Tennant, J.R. (1964) Evaluation of the Trypan Blue Technique for Determination of Cell Viability. Transplantation , 2, 685-694. https://doi.org/10.1097/00007890-196411000-00001
Crutchfield, A.L.M., Diller, K.R. and Brand, J.J. (1999) Cryopreservation of Chl a mydomonas reinhardtii (Chlorophyta). European Journal of Phycology , 34, 43-52. https://doi.org/10.1080/09670269910001736072
Orekhova, A., Barták, M. and Hájek, J. (2018) Post Rapid Freezing Growth of Antarctic Strain of Heterococcus sp. Monitored by Cell Viability and Chlorophyll Fluorescence. Cryobiology , 85, 39-46. https://doi.org/10.1016/j.cryobiol.2018.10.004
White, S., Anandraj, A. and Bux, F. (2011) PAM Fluorometry as a Tool to Assess Microalgal Nutrient Stress and Monitor Cellular Neutral Lipids. Bioresource Tec h nology , 102, 1675-1682. https://doi.org/10.1016/j.biortech.2010.09.097
Baker, N.R. (2008) Chlorophyll Fluorescence: A Probe of Photosynthesis in vivo . Annual Review of Plant Biology , 59, 89-113. https://doi.org/10.1146/annurev.arplant.59.032607.092759
Wellburn, A.R. (1994) The Spectral Determination of Chlorophylls a and b, As Well as Total Carotenoids, Using Various Solvents with Spectrophotometers of Different Resolution. Journal of Plant Physiology , 144, 307-313. https://doi.org/10.1016/S0176-1617(11)81192-2
Wowk, B. (2007) How Cryoprotectants Work. Alcor, Scottsdale. https://www.alcor.org/
Taylor, R. and Fletcher, R.L. (1999) Cryopreservation of Eukaryotic Algae—A Review of Methodologies. Journal of Applied Phycology , 10, 481-501. https://doi.org/10.1023/A:1008094622412
Voitsekhovskaja, O.V. and Tyutereva, E.V. (2015) Chlorophyll b in Angiosperms: Functions in Photosynthesis, Signaling and Ontogenetic Regulation. Journal of Plant Physiology , 189, 51-64. https://doi.org/10.1016/j.jplph.2015.09.013
Perrine, Z., Negi, S. and Sayre, R.T. (2012) Optimization of Photosynthetic Light Energy Utilization by Microalgae. Algal Research , 1, 134-142. https://doi.org/10.1016/j.algal.2012.07.002