Preserving microbial diversity has become a strategic undertaking. Thus, ex situ 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 Stichococcus bacillaris 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 Fv/Fm, 2.2 ± 0.25 g/g) to unpreserved cells and they are unsensitive to chosen agents, but the ratio between chlorophyll a and chlorophyll b 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 S. bacillaris cultures.
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