Viability of mass algal cultures preserved by freezing and freeze-drying

Cordero, Beatriz; Voltolina, Doménico

doi:10.1016/s0144-8609(97)00001-0

Cited by 31 publications

(21 citation statements)

References 11 publications

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“…Freezing with protective agents is another technique used in microalgal aquaculture applications when the main object is to maintain cell viability (Montaini et al 1995;Cordero and Voltolina 1997). Methanol has been reported to be the most suitable protective agent for the cryoprotection of certain cyanobacteria and algae (Hubálek 2003;Morris 1981).…”

Section: Discussionmentioning

confidence: 99%

Preservation methods of Tolypothrix tenuis for use as a cyanobacterial fertilizer

2006

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The suitability of the cyanobacterium Tolypothrix tenuis as a potential biofertilizer was evaluated by measuring cell viability in stored samples preserved by various methods, including freezing at −20°C, freeze-drying, desiccation as flakes and immobilization in alginate beads. The viability recovery and the retained viability index (RVI 10 ) were used as indicators of cell survival after 3 and 15 months storage. The highest recovery level (85%) was obtained by freeze-drying using skimmed milk as the resuspension solution for long-term storage. Dried alginate beads showed a better cell survival (RVI 10 0.25) than air-dried flakes (RVI 10 −0.63) after 15 months storage. However, desiccated flakes previously treated with Ca 2+ and Mg 2+ ions improved cell survival capacity at the longest period assayed (RVI 10 0.08), extending cell viability by 9 months compared to dried-powder material.

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Section: Discussionmentioning

confidence: 99%

Preservation methods of Tolypothrix tenuis for use as a cyanobacterial fertilizer

2006

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“…Low or no viability has been reported for the species C. gracilis (Cañavate & Lubian, 1995b), Chaetoceros sp. (Cordero & Voltolina, 1997), and Nannochloropsis oculata (Gwo et al, 2005) in the absence of CPA. Although viabilities up to 69% have been obtained without using CPA for Tetraselmis chuii, Nannochloropsis gaditana and Nannochloris atomus (Cañavate & Lubian, 1995b) and, certain Chlorococcaceae and Cyanobacteria (Day et al, 2000).…”

Section: Growth and Cryopreservationmentioning

confidence: 98%

“…In the case of C. calcitrans, its reaction to cryopreservation without DMSO could be explained by the excessive dehydration at the intracellular level caused by the formation of extracellular ice crystals that promote cell death (Tanaka et al, 2001;Day & Harding, 2007). Cordero and Voltolina (1997) in Chaetoceros sp., suggest that the presence of CPAs is necessary for post-freezing cell recovery in Chaetoceros microalgae as in other microorganisms. DMSO has shown high levels of toxicity in several species (Cañavate & Lubian, 1994;Fuller, 2004).…”

Section: Growth and Cryopreservationmentioning

confidence: 99%

“…In the case of species of the genera Chaetoceros, Chaetoceros gracilis (Cañavate & Lubian, 1995b, 1997a and Chaetoceros sp. (Cordero & Voltolina, 1997) have shown viabilities after cryopreservation lower than 40%. For the species Chaetoceros muelleri, cryopreservation has been successfully reported using DMSO 10% and 15%, however, there are not quantitative reports of viability (Rhodes et al, 2006).…”

Section: Introductionmentioning

confidence: 97%

“…For that reason, aquaculture facilities always look for strains of microalgae − from subcultures or cryopreserved collections − with biochemical profiles suitable for feeding commercial species (Cañavate & Lubian, 1995b;Cordero & Voltolina, 1997;Day & Harding, 2007).…”

Section: Introductionmentioning

confidence: 99%

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Cryopreservation of the microalgae Chaetoceros calcitrans (Paulsen): analysis of the effect of DMSO temperature and light regime during different equilibrium periods

Leiva

Dupré

2011

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ABSTRACT.We evaluated the effect of three variables (cryoprotectant temperature, light regime, and time of exposure to the cryoprotectant) throughout the equilibrium period during cryopreservation on the viability of the microalga Chaetoceros calcitrans (Paulsen). For this, the cryoprotectant dimethyl sulfoxide (DMSO) at 5% (v/v) was added at three different temperatures (4, 10, and 25ºC) before placing the microalgae in cryobiological straws for freezing. Once inside the cryobiological straws, the microalgal-cryoprotectant suspensions were subjected to the following light regimes for 15 or 45 min: complete light, complete darkness, light/darkness, and darkness/light. Suspensions were then frozen under controlled conditions and stored in liquid nitrogen. The viability index proposed by Cañavate & Lubian (1995b) was used to measure microalgal viability after cryopreservation. Results indicated that it is necessary to use a cryoprotectant to ensure the viable cryopreservation of C. calcitrans. Statistical analyses showed that the temperature of the DMSO influenced the viability of cryopreserved microalgae and that there was no synergistic effect between the other variables studied. The viabilities obtained with DMSO at 25ºC, 10ºC, and 4ºC were 34.9%, 27.8%, and 20.6%, respectively. Keywords: cryopreservation, diatoms, DMSO, temperature, light, equilibrium period.Criopreservación de las microalgas Chaetoceros calcitrans (Paulsen): análisis del efecto de la temperatura de DMSO y régimen de luz durante diferentes períodos de equilibrio RESUMEN. Se evaluó el efecto de tres variables − la temperatura del crioprotector, el régimen de luz y el tiempo de exposición al crioprotector durante el período de equilibrio en el proceso de criopreservación − sobre la viabilidad de la microalga Chaetoceros calcitrans (Paulsen). Para ello se utilizó, el crioprotector dimetilsulfóxido (DMSO) al 5% (v/v), el cual fue adicionado a tres temperaturas diferentes (4, 10 y 25ºC) antes de que las microalgas fueran introducidas dentro de pajuelas criobiológicas para su congelación. Una vez dentro de las pajuelas, las suspensiones de microalgas con crioprotector se sometieron durante 15 o 45 min a luz completa, oscuridad completa, luz/oscuridad y oscuridad/luz. Luego se congelaron de manera controlada y se almacenaron en nitrógeno líquido. La viabilidad de las microalgas posterior a la criopreservación fue medida a través del índice de viabilidad propuesto por Cañavate & Lubian (1995b). Los resultados indicaron que es necesario el uso de criprotector para la criopreservación viable de C. calcitrans. El análisis estadístico demostró que la temperatura del DMSO influye sobre la viabilidad de la microalga criopreservada y que no existe efecto sinérgico entre las demás variables de estudio. Las viabilidades alcanzadas usando DMSO a 25ºC, 10ºC y 4ºC fueron de 34,9%, 27,8% y 20,6% respectivamente.

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Comparative evaluation of different preservation methods for cyanobacterial strains

et al. 2012

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Viability of mass algal cultures preserved by freezing and freeze-drying

Cited by 31 publications

References 11 publications

Preservation methods of Tolypothrix tenuis for use as a cyanobacterial fertilizer

Preservation methods of Tolypothrix tenuis for use as a cyanobacterial fertilizer

Cryopreservation of the microalgae Chaetoceros calcitrans (Paulsen): analysis of the effect of DMSO temperature and light regime during different equilibrium periods

Comparative evaluation of different preservation methods for cyanobacterial strains

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