An interesting finding from the cold water strain collection project was that many species that predominate after the enrichment procedure were the same as the warm water species selected in previous collection efforts. The genera and species that were commonly found in both the cold water and warm water screening projects were C. muelleri, Amphora coffeiformis, Cyclotella, Navicula, and Nitzschia. However, some ochromonids and green coccoid algae were also isolated from the cold water collection effort; these types of alga were less commonly isolated during the warm-water selection procedures. Additional work would be needed to characterize these strains with respect to lipid production potential. Future work should look at Pubticeitfans acid profiles of oil found in the cold-water strains. Such cold-water i®assm? fn> o®fi? GUg h^, W .RIO tens atBraffds8)ttyrhaec m ^fo^li perform poorly as a feedstock for biodiesel because oLtheir low oxidative stability ecology of Tleurocnrysis carterae var dentata nov. (Prymnesiophyceae), a new and tendency to > polymerize >during combustion (Harrington et al. 1J3$p)i _>> , . coccolitnopnondifrom an inland3 saline pond in New Mexico, USA. Phycologica

¿JjSsgg. J.; Lemke, P.; Barclay, W.; Nagle, N. (1987) "Collection, screening, and characterization of lipid producing microalgae: Progress during Fiscal Year 1987." FY

1987 Aquatic Species Program Annual Report, Solar Energy Research Institute,

Golden, Colorado,' SERI/SP-231-32p6vnp. 27-42. , , ,

Johansen, J.R.; Theriot, E. (1987) "The relationship between valve diameter and number of central fultoportulae in Thalassiosira weissflogii (Bacillariophyceae)." J.

Phycost, 23MG'3-6Ji5lansen' J.R. (1988) "Physiological characterization of six lipid-producing diatoms from the southeastern United States." J. Phycol. 24:445-452.

Additional References:

Harrington, KJ. (1986) Biomass 9:1-17.

II.A.1.f. Development of a Rapid Screening Procedure for Growth and Lipid

Contentof Microalgae

By 1987, SERI researchers and subcontractors had collected approximately 3,000 algal strains. Most of these strains had not been well characterized, especially with respect to lipid production capabilities. As a consequence, work commenced on the development of a simple screening procedure to estimate the lipid contents of cells to determine which strains had the best potential as biofuel production organisms. Ideally, the procedure should be simple and reproducible so that it could be used as a standard method in numerous laboratories. The researchers hoped that such a screening tool would allow the size of the strain collection to be reduced to a manageable number (~200) representing the most promising strains.

Development of a rapid screen for lipid content.

In an attempt to develop a reproducible, easy-to-use screening procedure to identify algal strains with high lipid contents, Dr. Keith Cooksey (an ASP subcontractor at Montana State University) suggested that investigators explore the possibility of using the lipophilic dye Nile Red (9diethylamino-5H-benzo{a}phenoxazine-5-one) to stain cells. Nile Red was first isolated from Nile Blue by Greenspan et al. (1985), who showed that Nile Red will fluoresce in a nonpolar environment and could serve as a probe to detect nonpolar lipids in cells. Nile Red permeates all structures within a cell, but the characteristic yellow fluorescence (approximately 575 nm) only occurs when the dye is in a nonpolar environment, primarily neutral storage lipid droplets. Earlier work within the ASP by Dr. Steve Lien had shown the utility of Nile Blue in

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