Implications of a Catastrophic Refrigeration Failure on the Viability of Cryogenically Stored Samples.

Cryopreservation, the use of very low temperatures to preserve structurally intact living cells and tissues, is a key underpinning technology for life science research and medicine. It is employed to ensure the stability of critical biological resources including viruses, bacteria, protists, animal cell cultures, plants, reproductive materials and embryos. Fundamental to ensuring this stability is assuring stability of cryogenic storage temperatures.

Enabling large-scale production of algal oil in continuous output mode.

Large-scale algal oil production requires continuous outputs and a trade-off between growth and oil content. Two unrelated marine algae ( [CCAP 849/10] and [CCAP 211/21A]) that showed high oil production under batch culture were studied under controlled semicontinuous cultivation conditions. Three essential attributes maximized oil productivity: (i) downregulation of cell size to maximize light absorption under N limitation; (ii) low nutrient-depletion thresholds to trigger oil induction; (iii) a means of carbohydrate suppression in favor of oil.

Removal of metals from aqueous solutions using dried Cladophora parriaudii of varying biochemical composition.

Macroalgal biosorption has shown promise for the removal of metal ions from wastewaters, whose presence can pose a threat to the aquatic environment. There is a wealth of literature published on macroalgal biosorption, the common thread being that the biosorbent material was collected from the field, under undefined conditions. These studies offer little insight into the impact of prior cultivation or biomass production practices upon the biosorbent material, its adsorptive physico-chemical properties and its subsequent capacity for metal removal.

Effects of cryopreservation on viability and functional stability of an industrially relevant alga.

As algal biotechnology develops, there is an increasing requirement to conserve cultures without the cost, time and genetic stability implications of conventional serial transfers, including issues regarding potential loss by failure to regrow, contamination on transfer, mix up and/or errors in the documentation on transfer. Furthermore, it is crucial to ensure both viability and functionality are retained by stored stock-cultures. Low temperature storage, ranging from the use of domestic freezers to storage under liquid nitrogen, is widely being used, but the implication to stability and function rarely investigated.

Cryopreservation studies of an artificial co-culture between the cobalamin-requiring green alga and the bacterium .

Algal-bacterial co-cultures, rather than cultures of algae alone, are regarded as having the potential to enhance productivity and stability in industrial algal cultivation. As with other inocula in biotechnology, to avoid loss of production strains, it is important to develop preservation methods for the long-term storage of these cultures, and one of the most commonly used approaches is cryopreservation. However, whilst there are many reports of cryopreserved xenic algal cultures, little work has been reported on the intentional preservation of both algae and beneficial bacteria in xenic cultures.

Media Screening for Obtaining Haematococcus pluvialis Red Motile Macrozooids Rich in Astaxanthin and Fatty Acids.

Astaxanthin from is commercially produced in a two-stage process, involving green vegetative (macrozooid) and red aplanospore stages. This approach has been scaled up to an industrial process but constraints limit its commercial success and profitability, including: contamination issues, high pigment extraction costs, requirements for high light levels and photo-bleaching in the red stage. However, in addition to the aplanospore stage, this alga can produce astaxanthin in vegetative palmelloid and motile macrozooid cells.

A comparison of methods for the non-destructive fresh weight determination of filamentous algae for growth rate analysis and dry weight estimation.

The determination of rates of macroalgal growth and productivity via temporal fresh weight (FW) measurements is attractive, as it does not necessitate the sacrifice of biomass. However, there is no standardised method for FW analysis; this may lead to potential discrepancies when determining growth rates or productivity and make literature comparison problematic. This study systematically assessed a variety of lab-scale methods for macroalgal FW measurement for growth rate determination.

Bioremediation efficacy-comparison of nutrient removal from an anaerobic digest waste-based medium by an algal consortium before and after cryopreservation.

An algal consortium was isolated from an integrated steelmaking site at TATA Steel Strip Products Ltd. in Port Talbot, UK, and its bioremediation capacity tested. Excellent "bioremediation" was observed when the mixed culture was "applied" to diluted effluent from an enhanced anaerobic digestion plant at Dŵr Cymru Welsh Water at Port Talbot, UK.

Developing an ecologically relevant heterogeneous biofilm model for dental-unit waterlines.

This study monitored the biodiversity of microbes cultured from a heterogeneous biofilm which had formed on the lumen of a section of dental waterline tubing over a period of 910 days. By day 2 bacterial counts on the outlet-water showed that contamination of the system had occurred. After 14 days, a biofilm comparable to that of clinical waterlines, consisting of bacteria, fungi and amoebae had formed.

Challenges for the Maintenance and Cryopreservation of Multiple Isolates of Model Microorganisms: An Example Using the Marine Diatom Skeletonema marinoi.

Modern genomic and metabolomic tools have provided the possibility of generating and interrogating large datasets that can provide answers to previously imponderable taxonomic, evolutionary, ecological, and physiological questions. However, the curatorial tools needed to provide and maintain the relevant biological resources on which new knowledge can be built have not kept pace with this meteoric rise in scientific capacity, its associated activity, or the huge increase in published science. The availability of biological material of guaranteed identity and quality in Biological Resource Centers is fundamental for scientific research, but it crucially depends on there being adequate preservation/maintenance methods that are capable of ensuring phenotypic, genotypic, and functional security of the biological material(s).

Biofouling community composition across a range of environmental conditions and geographical locations suitable for floating marine renewable energy generation.

Knowledge of biofouling typical of marine structures is essential for engineers to define appropriate loading criteria in addition to informing other stakeholders about the ecological implications of creating novel artificial environments. There is a lack of information regarding biofouling community composition (including weight and density characteristics) on floating structures associated with future marine renewable energy generation technologies. A network of navigation buoys were identified across a range of geographical areas, environmental conditions (tidal flow speed, temperature and salinity), and deployment durations suitable for future developments.

Unlocking nature's treasure-chest: screening for oleaginous algae.

Micro-algae synthesize high levels of lipids, carbohydrates and proteins photoautotrophically, thus attracting considerable interest for the biotechnological production of fuels, environmental remediation, functional foods and nutraceuticals. Currently, only a few micro-algae species are grown commercially at large-scale, primarily for "health-foods" and pigments. For a range of potential products (fuel to pharma), high lipid productivity strains are required to mitigate the economic costs of mass culture.

Proteomic-based biotyping reveals hidden diversity within a microalgae culture collection: An example using Dunaliella.

Accurate and defendable taxonomic identification of microalgae strains is vital for culture collections, industry and academia; particularly when addressing issues of intellectual property. We demonstrate the remarkable effectiveness of Matrix Assisted Laser Desorption Ionisation Time of Flight Mass Spectrometry (MALDI-TOF-MS) biotyping to deliver rapid and accurate strain separation, even in situations where standard molecular tools prove ineffective. Highly distinctive MALDI spectra were obtained for thirty two biotechnologically interesting Dunaliella strains plus strains of Arthrospira, Chlorella, Isochrysis, Tetraselmis and a range of culturable co-occurring bacteria.

Comparison of screening methods for high-throughput determination of oil yields in micro-algal biofuel strains.

The phenotypic and phylogenetic diversity of micro-algae capable of accumulating triacylglycerols provides a challenge for the accurate determination of biotechnological potential. High-yielding strains are needed to improve economic viability and their compositional information is required for optimizing biodiesel properties. To facilitate a high-throughput screening programme, a very rapid direct-derivatization procedure capable of extracting lyophilized material for GC analysis was compared with a scaled-down Folch-based method.

Post-cryopreservation viability of the benthic freshwater diatom Planothidium frequentissimum depends on light levels.

Over recent years, several planktonic and benthic freshwater diatom taxa have been established as laboratory model strains. In common with most freshwater diatoms the pennate diatom Planothidium frequentissimum suffers irreversible cell shrinkage on prolonged maintenance by serial transfers, without induction of the sexual cycle. Therefore, alternative strategies are required for the long-term maintenance of this strain.

Isolation of clonal cultures of endosymbiotic green algae from their ciliate hosts.

Using Paramecium bursaria as a model organism improved protocols have been developed to isolate clonal endosymbiotic algae. This involved micromanipulation of individual protists, rupturing to release endosymbionts followed by enrichment on complex media and a series of plating steps, under low light (PAR ~10μmol photons m(-2)s(-1)).

Effects of temperature and nutrient regimes on biomass and lipid production by six oleaginous microalgae in batch culture employing a two-phase cultivation strategy.

Commercial success of algal-based biofuels depends on growth characteristics and lipid metabolism of the production species. The oleaginous microalgae, Thalassiosira pseudonana, Odontella aurita, Nannochloropsis oculata, Isochrysis galbana, Chromulina ochromonoides, and Dunaliella tertiolecta, were cultivated under a matrix of two temperatures (10 and 20 °C) and two nutrient regimes (deplete and replete). For all species, a strong negative correlation between growth rate and lipid content was observed.

Overcoming biological constraints to enable the exploitation of microalgae for biofuels.

Microalgae have significant potential to form the basis of the next biofuel revolution. They have high growth and solar energy conversion rates. Furthermore, their osmotolerance, metabolic diversity and capacity to produce large amounts of lipids have attracted considerable interest.

Encapsulation dehydration colligative cryoprotective strategies and amplified fragment length polymorphism markers to verify the identity and genetic stability of euglenoids following cryopreservation.

An encapsulation/dehydration procedure was developed for Euglena gracilis Klebs as a 'model alga' to examine various cryoprotective regimes combined with controlled rate cooling to cryopreserve other Euglenoid taxa. Cryoprotective variables were optimised to enable reproducible growth following a combination of alginate encapsulation, sucrose osmotic dehydration, air desiccation, methanol treatment, cooling to -40 degrees C and plunging into liquid nitrogen (LN). Amplified Fragment Length Polymorphism (AFLP) analysis was adapted to: (i) verify algal identity by discriminating between different Euglenoids and (ii) examine the genetic stability of algal cultures prior to various stages of cryoprotective treatments and following exposure to LN.

Tolerance of the resting cysts of Colpoda inflata (Ciliophora, Colpodea) and Meseres corlissi (Ciliophora, Spirotrichea) to desiccation and freezing.

The survival of ciliate resting cysts, in the presence and absence of soil, was studied under two environmental stresses: desiccation and freezing. Laboratory strains of the common species Colpoda inflata and the rare species Meseres corlissi were used in these experiments, which yielded the following results: 1) Freezing of cysts in soil with a residual moisture level exceeding approximately 30% was destructive for both species. 2) Survival of Meseres corlissi cysts depended largely on the presence of soil.

Cryopreservation of resting cysts of the freshwater ciliate Meseres corlissi by conventional two-step methods and one-step vitrification protocols.

Trophic cells of the freshwater ciliate Meseres corlissi CCAP 1647/1 proved to be recalcitrant to all the cryopreservation methods tested; however, resting cysts of this strain were amenable to both conventional two-step cryopreservation and ultra-rapid vitrification methods. Conventional controlled rate cooling and Mr. Frosty cooling methods, employing 5 percent dimethylsulphoxide (DMSO) as a cryoprotectant were effective in preserving cysts in either liquid medium or soil suspensions.

Biobanking.

Biobanks, more formally known as biological resource centers (BRCs), form an "unsung" yet critical component of the infrastructures for scientific research, industry and conservation, without which much of the current scientific activity involving microbial cultures and cell-lines would be effectively impossible. BRCs are de facto depositories of "biological standards" holding taxonomic and other reference strains on which much of the associated published science and industrial standards are built and upon which some significant international commercial and ethical issues rely. The establishment and maintenance of BRCs is a knowledge- and skill-rich activity that in particular requires careful attention to the implementation of reliable preservation technologies and appropriate quality assurance to ensure that recovered cultures and other biological materials perform in the same way as the originally isolated culture or material.