Culturing Microalgae at the Small Culturing Microalgae at the Small Scale . Culturing Microalgae at

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  • Presentation to ABO 7th Annual Algae Biomass Summit, Orlando, October 2, 2013.

    Willie Wilson, Ph.D.

    Director The Provasoli-Guillard National Center for

    Marine Algae and Microbiota (NCMA)

    Bigelow Laboratory for Ocean Sciences

    Culturing Microalgae at the Small Scale

  • Culturing Microalgae at the Small Scale

    A Case of Perpetual Care. Why cultivate (domesticate) algae?

    What are your objectives?

    Does the need justify the effort?

    Are there other means of doing what you need?

    What facilities are needed?

    A defined area of controlled temperature to grow cultures.

    Facilities for making up media.

    Means of sterilizing the media?

    What type of media do you need?

  • Simplest to the most complex!

    Temperature/light regulated growth chamber

    Laboratory North Facing Window

    The major control is temparature.

    Light intensity and duration is next.

    Alarms or warning controls.

  • Maintenance in tube culture –

    Small batch mode

    30-40 mL of culture

    Maintained with frequent transfers – ideal is monthly intervals

    Needs:

    Sterile glass culture tubes

    Sterile medium

    Appropriate growing space

    Handling materials – glass pipettes heat or autoclaved.

    Laminar flow hood

    Bacteriological sterile technique needed!

    New curators if the collection becomes of any size.

  • Media making do’s and dont’s

    Defined media - can be made in specified limits the same each time

    many species will not grow as well as in other media

    costly since base water is usually highly purified

    excellent for physiology or where conditions need to be defined within narrow limits

    Examples: Wilson’s NH15 and Aquil

    Enriched sea water media (ESW)

    requires good quality, off shore seawater. Great if you live near the Gulf of Mexico, Sargasso Sea or Eastern Mediterranean

    Besides having a mode of acquiring the sea water (ships, pumping systems, cold dark storage) you must be aware of seasonal variations.

  • Example of successful sea water enriched media.

    Oldest and a stabile stand by – Erdschribers Media

    Problem – soil extract plus N and P

    Advantage – grows nearly everything (Ask Richard and Maria)

    The Provasoli variations –

    Sea water enrich with trace metals tailored to the species

    Precipitation often a problem

    The Guillard Series – beginning with f/2, K medium, L1 medium et al.

    With understanding the need for selenium, many of the problem species have been cultivated. Open ocean species are no longer a problem.

  • Work stations.

  • Think about your culture space, walk-in incubators are convenient, and flexible.

  • Reach-in incubators work well. Note: light level variability. Different culture vessels. Ways to label growth medium.

  • Accurate labeling is paramount.

  • Seaweeds are algae too! ..and they are easy to culture.

  • Cryopreservation save collection managers a small fortune…..

  • A role for bioresource centers (BRCs) in the 21st century.

    • A Bioresource Center • Mission and Vision • Microbial (Algal) Commodities • An NCMA Snapshot • Leading an Algal Industry

    Cluster in Maine Practicing the ART OF POSSIBILITY

  • Time to move???

  • NCMA’s status is a service center of Bigelow Laboratory for Ocean Sciences, a non-profit 501(c)(3) organization. Income derived from federal grants, culture fees and services support the functions of our mission. NCMA’s mission is to serve as a central bioresource center whose core activity is to receive, maintain and distribute living cultures of marine and freshwater algae, bacteria and viruses; to provide technical expertise and services; and to provide educational resources for culture isolation and curation to scientists, educators, biomedical researchers, and businesses worldwide. NCMA’s vision is to become the global leader in the provision of algae, protozoa, bacteria and viruses, together with related products for academic research and commercial ventures. By 2020 the NCMA brand will be widely recognized for first class customer service and intellectual capability.

  • Microbial ‘Commodities’ Oceanic microbiota

    (protists, algae,bacteria, archeae, viruses) are the

    largest untapped biotechnological resource

    on the planet.

    Glynn Gorick

  • The NCMA holds representatives from 39 Classes of Algae (all the major photosynthetic groups)

    Apicomplexa Aurearenophyceae Bacillariophyceae Bicosoecophyceae

    Ciliatea Charophyceae Chlorarachniophyceae

    Chlorophyceae Chromerida

    Chromophyte

    Chrysomerophyceae Chrysophyceae

    Bolidophyceae

    Coscinodiscophyceae

    Cryptophyceae

    Cyanophyceae

    Dictyochophyceae

    Dinophyceae

    Euglenophyceae

    Eustigmatophyceae Fragilariophyceae

    Glaucophyceae Pelagophyceae

    Phaeophyceae

    Phaeothamniophyceae Pinguiophyceae

    Porphyridiophyceae

    Prasinophyceae

    Prymnesiophyceae

    Raphidophyceae

    Rhodophyceae

    Schizocladiophyceae Synchromophyceae

    Synurophyceae Trebouxiophyceae

    Ulvophyceae undetermined

    Apicomplexa

    Aurearenophyceae

    Bacillariophyceae

    Bicosoecophyceae

    Bolidophyceae

    Charophyceae

    Chlorarachniophyceae

    Chlorophyceae

    Chromerida

    Chromophyte

    Chrysomerophyceae

    Chrysophyceae

    Ciliatea

    Coscinodiscophyceae

    Cryptophyceae

    Cyanophyceae

    Dictyochophyceae

    Dinophyceae

    Euglenophyceae

    Eustigmatophyceae

  • Geographical Location of Strains

  • NCMA: A Snapshot

    • The world’s largest and most diverse living archive of marine microalgae.

    • 35 years in the algae business. • Started as a algal seed stock for the aquaculture industry. • A repository for public and private collections of algae. • 2,720 strains of marine, brackish, hypersaline and freshwater

    algae; including cyanobacteria and macroalgae. • 359 genera and 723 species. • 6 products available for every strain. • 5 growth temperatures (polar to tropical). • On-site & off-site back-up and cryopreservation. • Expansion to include new Bacteria and Virus collections. • New web site to enhance customer experience (Oct 2013).

  • Products and Services

    • Starter cultures • Nucleic acids from algae • Culturing Techniques Courses • Private collections • International Depository Authority (patent depository) • got algae? T-shirts! • Research services • Isolations/clean up/taxonomic ID • Services through our partners (e.g. powdered algae) • Other service centers at Bigelow

    – Single Cell Genomics – Flow Cytometry – Analytical Services – Seawater Facility

  • VISION To create a complete algal supply chain; from seed cultures to biomass infrastructure, processing, testing, and development of market opportunities; that will allow us to lay a foundation to stimulate algal economic development, and make Maine one of the top states in the U.S. to run an algal-based business. It will be a process driven by innovation, education, collaboration, and branding.

    Proposal for an Algal Industry Cluster in Maine

  • NCMA Entrepreneurship The Future is Bright!

    Because the oceans of the world represent a vast frontier of undiscovered compounds and organisms, the prospect exists for the NCMA to work with numerous private industries to explore commercial opportunities for new products and technologies in a variety of commercial markets.

  • Innovation Gets You Recognized

  • The NCMA Team

  • Questions

    Dr. Willie Wilson

    wwilson@bigelow.org https//ncma.bigelow.org

    207-315-2567 (x310)

    Slide Number 1 Slide Number 2 Slide Number 3 Slide Number 4 Slide Number 5 Slide Number 6 Slide Number 7 Slide Number 8 Slide Number 9 Slide Number 10 Slide Number 11 Slide Number 12 A role for bioresource centers (BRCs) in the 21st century. Slide Number 14 Slide Number 15 Slide Number 16 Slide Number 17 Slide Number 18 Slide Number 19 Slide Number 20 Slide Number 21 Slide Number 22 Slide Number 23 The NCMA holds representatives from 39 Classes of Algae� (all the major photosynthetic groups) Geographical Location of Strains NCMA: A Snapshot� Slide Number 27 Slide Number 28 Products and Services� Slide Number 30 NCMA Entrepreneurship�The Future is Bright! Slide Number 32 Slide Number 33 Questions